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* ''WesternAnimation/AmericanDad'': Sanders, Clifford, Clown with Glasses, Hercules, Miriam, and Duper (he was replaced with a clone) have all died permanently.
* B'wana Beast, [[ComicBook/BlueBeetle Ted Kord]], the original ComicBook/BlackCanary, and the entire ComicBook/DoomPatrol are all killed off during the three season run of ''WesternAnimation/BatmanTheBraveAndTheBold''. The deceased heroes do appear in the show's GrandFinale, but only as part of a [[BreakingTheFourthWall Fourth Wall-breaking]] wrap party that [[BittersweetEnding shows the entire cast together one last time before everything fades to black]].
* ''WesternAnimation/TheBoondocks'' Season 1: "Granddad's Fight." The karmic fate of life-long, Five-Star MagnificentBastard Colonel H. Stinkmeaner in an ironic CurbStompBattle with Robert Freeman: Stinkmeaner had beaten Robert in an argument over a parking spot, and the entire episode revolves around Huey making him out to be a version of a blind assassin, and Robert training for a rematch. It turns out Stinkmeaner was just a crotchety old man who got lucky. Unfortunately, he's such a temporal {{Jerkass}}, absolutely ''nobody'' realizes this until Robert pulverizes him. The beating is so bad, Stinkmeaner ends up stone-cold dead with a solid punch to the face. Feeling remorseful at his grave, Robert prays to God for forgiveness at the end of the episode.
** Although dead, Stinkmeaner [[TookALevelInBadass returns]] in Season 2: "Stinkmeaner Strikes Back". Stinkmeaner has been sent to Hell after death, but now proves he can seriously fight back- by pummeling his way through every demon who steps up to challenge him. Satan himself is so marveled at Stinkmeaner's exposition, he sends him back to Earth to [[ResurrectedForAJob spread ignorance and chaos among the black community]], also giving him the opportunity to seek revenge against the Freeman family. As a ghost, he posesses Tom Dubois and trashes most of his house. After Uncle Ruckus and Riley fail miserably at exorcising Stinkmeaner, Ghostface Killah's spirit shows up and clues Huey in to the solution: put Stinkmeaner at peace by having Ruckus spout off about how he hates black people, a hate Stinkmeaner firmly shares with him. Moments later, he's exorcised and sent right back to hell.
** In yet another Stinkmeaner-related episode, death strikes again, this time nailing Bushido Brown, via [[OffWithHisHead on-screen decapitation]] during his fight in Season 3: Stinkmeaner 3: The Hateocracy". The Colonel apparently palled around with a group of disgruntled elderly[[{{Jerkass}} jerks]] who formed the Hateocracy, a gang that wreaked havoc within the walls of a nursing home. The members of the gang show up to [[CurbStompBattle curb-stomp]] the Freeman family in what looks like a RoaringRampageOfRevenge, so the Freemans hire Bushido as their equally jerkish bodyguard, who actually holds off the Hateocracy- until he catches a [[EverythingsBetterWithSpinning spinning disk of doom to the neck]]. The Freeman family's reaction? A group [[PrecisionFStrike "OH, SHIIIIT!"]].
*** It gets more bizarre when Stinkmeaner's name comes up. Robert tries to reason with the Hateocracy to leave them alone and forget revenge, and they reveal they couldn't care less about the deceased Colonel- [[ForTheEvulz they're just ruining lives for the hell of it]]. Luckily, an army of cops shows up in the nick of time to cart off the unruly trio. Taking a jab of his own at his ungrateful cohorts, Stinkmeaner's ghost, who has been narrating on and off throughout the episode, [[LampshadeHanging hangs a cynical lampshade on his fate over his old pals]]: He thinks death and hell are ''better'' than being in jail!
** In the final part, "Stinkmeaner: Begun the Clone War Has" in Season 4, a clone of Stinkmeaner begins picking on Robert and puts him through a HumiliationConga. [[CorruptCorporateExecutive Ed Wuncler Sr.]] then forces Robert to fight him again so he can make money off it, which culminates in Robert delivering a NoHoldsBarredBeatdown and telling Stinkmeaner to leave him alone. He agrees, because he's just a clone with none of Stinkmeaner's memories and barely even knows Robert.
* Don King in ''WesternAnimation/CelebrityDeathmatch'' normally comes back from the dead after getting killed in the audience, usually on accident or by a monster. When he actually fought in a match, Donald Trump killed him, only to never be seen again.
** Stacy Cornbred dies after spontaneously combusting in "The Prophecy". She came back as a zombie in one Halloween episode, but was killed again when Debbie Matenopoulos shoves her through the side of a door, shredding her into bits. She stays dead from that point on, even though Johnny and Nick could easily have revived her with the Time Machine both times.
* The GrandFinale of ''WesternAnimation/CodenameKidsNextDoor'' implied this happened to The Delightful Children.
* The series ''WesternAnimation/DarkwingDuck'' has two villains who die (on screen): Splatter Phoenix, who melts while she gives a rambling speech as she's erased from existence, and Mayor Synapse who turns into a powerful almost indestructible being until the main heroes get him to blast and explode "graphically".
* In an aversion of [[JokerImmunity the very trope named after him]], the Joker eventually got this treatment in the Franchise/{{DCAU}}. Yes, having proven himself as much of a survivor as his counterpart in the comics, every bit the "no one" in NoOneCouldSurviveThat, ''WesternAnimation/BatmanBeyondReturnOfTheJoker'' took the big leap and put him down once and for all - killing him twice; first his body was destroyed and buried deep within Arkham Asylum in flashback, then the chip containing a copy of his DNA and memories was burned out, just to make sure he won't be coming back. (In just a bit of a cheat, the event occurs at the far end of the universe-at-large's history, allowing him to show up in stories taking place earlier; ''WesternAnimation/JusticeLeague'' took advantage of this.)
* ''WesternAnimation/DefendersOfTheEarth'' features several onscreen deaths. The first character to die in the series is Flash Gordon's wife (who may or may not be Dale Arden) whose life force is drained by Ming's Inquisitor as Ming tries to force her to reveal Flash's whereabouts. In her case, however, the trope is zigzagged; though she stays dead, her son, Rick, uses a crystal containing her life essence as the personality base for the Defenders' supercomputer, Dynak-X.
* In ''WesternAnimation/FamilyGuy'' a few recurring characters such as Mr. Weed, Paddy Tanniger, Vern and Johnny, and Francis Griffin have been killed off. Despite NegativeContinuity, any returns are made by their ghosts.
** Joan Quagmire died on a technicality, having tried to grab Death's hand. As she was married to the "deceased" Glenn Quagmire and having taken his last name, he agreed to take her instead. Being a bit off didn't hurt, either....
** It was revealed in a Season 6 episode that Joe's son Kevin had died in Iraq. This was done because the writers thought he was boring. However, [[SubvertedTrope he came back in the recent Thanksgiving episode (it turned out he had faked his death)]], unfortunately there was no time left for a SpotTheImposter moment, so there was NoEnding.
** Stewie killed his half-brother Bertram in 15th-century Italy, which creates a very odd temporal paradox...then came the videogame ''VideoGame/FamilyGuyBackToTheMultiverse''
** In fact, [[ButtMonkey Meg]] and [[{{Jerkass}} Peter]] are the only characters to die and come back to life. Later, Stewie, Chris, and Mort have been killed but only as a gag. Brian also died but was revived by Stewie.
** In the season 9 premiere, [[TonightSomeoneDies the trailers advertised that someone would die]]. As it turns out, Muriel Goldman, Derek, Diane Simmons, and onetime characters Stephanie and Priscilla were killed off.
*** Amusingly subverted with [[AdamWesting James Woods]]. He's seemingly killed off for good in that same episode only to turn up later on (complete with deliberately hilarious and absurd VoodooShark explanation for his return to boot).
** [[BastardBoyfriend Jeff]] in "Screams of Silence".
** Horace dies in "Save the Clam".
** Mental Mike in the Season 12 episode "A Fistful of Meg".
** Death also dies in his last appearance.
* In the ''WesternAnimation/{{Futurama}}'' episode "The Thief of Baghead", over-the-top robot actor Calculon dies when he ingests food colouring (which is deadly to robots) in his enactment of Romeo's death in ''Theatre/RomeoAndJuliet'' to win an acting competition ([[ShootTheShaggyDog he lost]]). His death receives little attention by the cast and Calculon is revived and killed again in "Calculon 2.0" to receive a send-off episode.
* Webstor has the distinction of being the only member of the regular cast to truly die in the ''Franchise/MastersOfTheUniverse''. In the "Web of Evil" episode of ''WesternAnimation/HeManAndTheMastersOfTheUniverse2002'', he was assaulted off-screen by the Snake-Men as the heroes were closing in, then brutally beaten by them and collapsing dead on the ground when the good guys finally reached his lair. However, the scene depicted his death in an ambiguous manner, so fans were left to debate themselves whether or not he had survived until after the show's cancellation, when [[WordOfGod Ian Richter]] confirmed that he did not. (It is entirely possible that the writers had planned for the possibility to reverse this decision and include him in a later story, seeing as the series was cancelled three episodes later.)
* In ''WesternAnimation/JonnyQuestTheRealAdventures'', while the show's other main villains tended to get the NoOneCouldSurviveThat treatment in their last episodes -- and since the show didn't get a third season, it's probably safe to say that they're gone for real -- Ezekiel Rage had an established tendency to cheat death. Therefore, [[DroppedABridgeOnHim just to be sure]], he got sent back to prehistoric times and blown up by a nuclear bomb, [[NeverFoundTheBody leaving a skull behind]] for good measure. DeaderThanDead? Deader than dead.
* ''[[WesternAnimation/JusticeLeague Justice League Unlimited]]'' killed off most of the LegionOfDoom via an exploding spaceship in "Alive", the second-to-last episode of the series. While a small number were explicitly shown to survive, a number of minor villains like the Shade, Silver Banshee, Rampage, Neutron, and many others were presumed dead. Lex Luthor and Darkseid are apparently killed in the finale, "Destroyer", but Superman doubts that Lex is truly gone. [[WordOfGod Dwayne McDuffie]] has stated that Luthor and Darkseid now form part of the Source Wall, meaning they're technically alive, but unlikely to ever trouble the League again.
* Warhok and Warmonga from ''WesternAnimation/KimPossible'' were confirmed by WordOfGod to be dead for real after Ron used his Mystical Monkey Powers to throw them into their own ship after they threatened to kill Kim, after which the ship exploded. They're about the only villains in the series to be killed for real... Except perhaps for [[TakenForGranite Monkey Fist]].
* In ''WesternAnimation/KingOfTheHill'', quite a few recurring or main characters have been killed off over the years including Luann's first boyfriend Buckley, Buck Strickland's mistress Debbie, Pops Poppaseto, Cotton's war buddy Topsy, and most recently Hank Hill's father Cotton Hill.
* In the ''WesternAnimation/MyLittlePonyFriendshipIsMagic'' Season 5 episode "The Lost Treasure of Griffonstone", Arimaspi is shown to have fallen into the Abysmal Abyss, and his skull is later seen.
** G1 had Lavan, who died in similar way to Sombra (although a little less graphically), after his magic beam ricocheted from the princess ponies' magic wands back at him.
** And before that, the G1 TV special ''Rescue at Midnight Castle'' that kicked off the original show ended with Tirek's defeat via a giant rainbow-colored tornado, much like how Luna was stripped of [[SuperpoweredEvilSide her Nightmare Moon persona]] [[MythologyGag in the second episode of]] ''Friendship is Magic''. Except that Tirek wasn't redeemed by it, but killed outright.
*** Tirek and Lavan's deaths are notable not only because a series based on something as innocent as the ''My Little Pony'' series is probably the last place anyone would expect to see someone die, but also because killing off characters was something very rare to be seen in ''any'' TV cartoon in the 1980s.
* In ''WesternAnimation/ThePowerpuffGirls'', this possibility arises with Dick Hardly, a one-episode villain of "Knock It Off", who, within the episode, cemented himself as probably one of the most evil villains on the show, may actually have been the only human villain to ''die'', considering that, after his episode, him and his various PPG knockoffs ([[HoistByHisOwnPetard which were the ones that did him in]]) were ''never'' seen again.
** Bunny in "Twisted Sister." The girls attempt to create a fourth Powerpuff Girl using substitutes for sugar and spice, and their subjective forms of everything nice, as well as an "accidental" dose of Chemical X. The product is an apparently mentally disabled sibling with a black and white sense of right and wrong, which causes her to not quite get the hang of crime fighting. The girls, once they find out what trouble she has been up to, tell her she did bad and cause her to flee in tears. The criminals that Bunny released as a result of [[ItMakesSenseinContext viewing the police as bad in her black and white view of the world]], attack and overwhelm the girls, and Bunny hears their cries and comes back to save them. The ensuing battle causes her already unstable form to reach its maximum capacity, [[TearJerker and she explodes after proudly proclaiming that she finally did good.]] The narrator confirms she's not coming back.
-->'''Narrator:''' (tearfully) And for the first, and final time, the day is saved thanks to Powerpuff Bunny.
* Parodied in an episode of ''WesternAnimation/TheSimpsons''. Moe gets a facelift and becomes handsome, and is awarded a role on a soap opera. After reading a new script that says he dies, he retaliates by revealing all of the major plot twists in the show for the next year on air (the show was filmed live, apparently). The director than angrily informs him that his character wasn't being killed off and that that page of the script was a dream sequence.
** And not parodied with the very "real" (at least in animation terms) deaths of Bleeding Gums Murphy and Maude Flanders. Dr. Marvin Monroe was also apparently dead enough to have a hospital named in his memory (but [[UnexplainedRecovery he got better...]]).
** Maude Flanders counts as this, and in a TonightSomeoneDies fashion no less.
** When the hated [[ShooOutTheNewGuy Poochie]] is killed off, Krusty signs a legal document swearing that he will never, ever come back. And then he came back in a later Itchy and Scratchy episode as a [[TheCameo cameo]] guest to one of Scratchy's funerals.
** This trope finally caught up with Mona Simpson, Homer's mother. What makes this death particularly impacting is her successive NotQuiteDead track record in previous episodes.
** Mob boss Fat Tony dies of a heart attack, but in a SubvertedTrope, he was replaced by his cousin Fit Tony, who got so fat he looked just like Fat Tony, then changed his name to Fat Tony.
** Frank "Grimey" Grimes is dead and his son confirmed it.
** Possibly averted with Lionel Hutz's habit of repeatedly running over Judge Snyder's son. Being repeatedly run over might be enough to kill the Judge's son, but like Mr. Burns in "Who Shot Mr. Burns? Part 2", [[UnexplainedRecovery his conditions must have been upgraded to alive at the "better" Springfield General Hospital everytime.]] Because Lionel Hutz, like Troy [=McClure=], was retired when voice actor Creator/PhilHartman was killed off for real by his wife in a murder-suicide, it is not known of Hutz's habit has continued and the Judge's son is always killed, then revived.
** Due to her voice actress Marcia Wallace's death, WordOfGod is that Edna Krabappel has become this. After Marcia's death Edna was given a few silent cameos up until the end of Season 25 when Nelson and Ned Flanders are seen mourning at her grave.
* ''WesternAnimation/SouthPark'':
** Unless Isaac Hayes is somehow reanimated in real life, Chef is likely gone for good, despite his "Darth Chef" remaking.
** Inverted with [[TheyKilledKennyAgain Kenny]]. He was killed off for real, but returned at the end of the next season, with no explanation given. [[note]]Originally, Kenny ''was'' supposed to stay dead, since [[Creator/TreyParkerAndMattStone Parker and Stone]] were getting tired of killing him off in every episode and were running out of original ways to do so. Butters was meant to take his place as the new ButtMonkey character, but meddling from Creator/ComedyCentral meant the duo had to change these plans. [[/note]]
** Other recurring characters to have died over the years include Ms. Choksondik, Ms. Crabtree, the mayor's aide named Ted, and most recently Pip.
* ''WesternAnimation/StarVsTheForcesOfEvil'':
** While a few of Ludo's minions managed to escape the destruction of his castle at the end of "Storm the Castle", most of them were killed in the blast.
** Lekmet of the Magic High Commision carks it when he uses his life to heal Moon in "Starcrushed". To solidify the fact that he's dead, his body turns to ash.
** [[BigBad Toffee]] bites it by the end of "The Battle For Mewni", via WaveMotionGun and being crushed. While [[HealingFactor he could regenerate from any injury]] and had previously been NotQuiteDead, what really nails his fate is the fact that Moon's deal with Queen Eclipsa in the past meant the latter would be freed from her crystal prison once Moon's enemy (Toffee) was dead. At the end, said crystal begins to crack by itself.
* ''WesternAnimation/StarWarsTheCloneWars'' mostly plays it straight. Of course, due to being DoomedByCanon, none of the characters who appeared in ''Film/RevengeOfTheSith'' or the other movies taking place after it can be Killed Off for Real in the show, but no one else is a hundred percent safe.
* ''WesternAnimation/StarWarsRebels'': In [[Recap/StarWarsRebelsS3E18TwinSuns "Twin Suns"]], Darth Maul is finally killed by Obi-Wan Kenobi on Tatooine. The official ''Star Wars'' website even put up a ReallyDeadMontage just to confirm to fans that no, he's ''not'' coming back this time.
* At the end of the ''WesternAnimation/SupermanTheAnimatedSeries'' two-parter "Apokolips Now", after Darkseid's invasion of Earth is thwarted and just before he leaves he murders Superman's friend Metropolis Police Inspector Dan Turpin with his Omega Beams in front of Superman and a crowd of onlookers. Some fans hoped Turpin was just teleported to Apokolips like Kalibak had been after ''he'' got apparently zapped, but it seems he's gone for good.
* ''Franchise/{{Transformers}}'' as a whole has never shyed away from this.
** ''WesternAnimation/BeastWars'' continually killed its cast, despite the fact that ''Beast Wars'' characters are a bit hard to kill, as robots they have survived deadly attacks and being blown to bits (in which case they can easily be repaired). Some simply died, in one case to make room for new characters. A surprising number died, were brought back, and died again. Two of them died, were brought back as a fused form of the two, and then died. And two of them died [[ChekhovsGun with the possibility that they might return]], but didn't.
*** Major characters to permanently die are (in order of death) Terrorsaur, Scorponok, Dinobot, Tarantulas, Rampage, Depthcharge, Tigatron, Airazor (both as Tigerhawk), Inferno, Quickstrike, and Dinobot II. Major characters to survive to the end of the final episode are Optimus Primal, Rattrap, Cheetor, Rhinox, Blackarachnia, Silverbolt, Megatron, and [[JokerImmunity Waspinator]].
*** If you count the comics, the list of permanently dead characters is reduced to both Dinobots, Rampage, Tigerhawk, Inferno and Quickstrike.
*** And the in the sequel series, ''WesternAnimation/BeastMachines'', Optimus, Megatron, and Rhinox are killed as well. Leaving the only six characters alive at the end to be Rattrap, Cheetor, Blackarachnia, Waspinator, Nightscream, and Botanica.
** In ''WesternAnimation/TransformersAnimated'' Blurr is killed, pretty graphically, for a machine. Namely he was crushed into a cube and dumped down a trash chute.
*** The finale also killed off Prowl and Starscream. Prowl sacrificed his Spark to save the city, and Starscream's [=AllSpark=] fragment, the only thing keeping him alive after he was killed by Megatron in "Megatron Rising", was pulled out of his head. To hammer the point home, their bodies turned completely gray, the telltale sign in this series that a Transformer is really dead barring interference from the [=AllSpark=] itself.
** Jazz was killed by Megatron in the final battle of the first live-action movie. The Fallen was killed by Optimus at the end of the sequel and every Decepticon (who isn't Megatron) that has been killed has stayed dead so far.
** ''WesternAnimation/TransformersPrime'' killed off Cliffjumper in the first episode, then brought him [[CameBackWrong back as a zombie]] in the second episode only to die ''again''. Skyquake and Makeshift were also killed in their debut episodes, the former became another zombie while the latter is staying dead, [[TooPowerfulToLive because he was too powerful to stay alive]]. WordOfGod is that "when we kill a character, ''we kill a character''."
*** "Crossfire" has added Breakdown to the scrap heap.
*** "Hurt" added Hardshell.
*** Dreadwing in "Regeneration" - killed by Megatron after refusing to follow orders to ''not'' execute Starscream who zombified his deceased brother Skyquake. Lovely.
* ''WesternAnimation/TheVentureBrothers'' has quite a few, most [[TearJerker heartbreakingly]] 24's death by exploding muscle car.
* In ''WesternAnimation/WinxClub'' they have killed off the main villains of season 2, 3 and 4 at the end of those seasons. In addition to this, we lost [[HeroicSacrifice Nabu]] in season 4 (though the Cinelume version implies that he is alive by saying he might wake up and having a small cameo at the end of the season finale, however, his death is confirmed in the Nickelodeon version). Nabu was about to be revived by the Black Gift, but Ogron steals it and wastes it on a wilting flower. And apparently, [[ForgotAboutHisPowers Bloom never thought of using her healing powers to at least try to nurse Nabu back to health.]]
* ''WesternAnimation/{{WITCH}}'' killed off quite a few of the lesser villains, such as Tridart and Ember being disposed of by Nerissa when they were no longer needed. In the finale, Tracker is also subsequently impaled and killed.
* ''WesternAnimation/YoungJustice'':
** Kent Nelson a.k.a. ComicBook/DoctorFate. His spirit did reside in the Helmet of Fate for a while, but Fate eventually got tired of his nagging every time someone used the helmet and sent him off to the afterlife.
** In the second season SeriesFauxnale, Kid Flash is disintegrated when he absorbs too much energy helping The Flash and Impulse get rid of an energy field that threatened to envelop the Earth. To honor his memory Impulse [[TakingUpTheMantle takes up his mantle]], and to hammer the point home he gets a hologram in the garden like Jason Todd, Tula and Ted Kord.