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Lucy BakerThe main protagonist and the narrator. She's a DC (detective constable) who works under Inspector Layton.
- Curtains Match the Window
- Declarative Finger: One of her frequent sprites is this.
- Expy: Of Luke Triton from the original Professor Layton series. They even wear similar-looking hats.
- Naïve Newcomer: She's start out at the beginning of the game as a new officer starting out as Alfendi's assistant.
- Nice Hat
- Player Character: Despite the title Layton Brothers, she's the character you actually play as.
- Plucky Girl: Lampshaded by the "other" Alfendi in case 3.
- Pungeon Master: Has a tendency to be this. This is taken Up to Eleven in Case 8 however, as it dealt with animals and many of her responses were puns about the people/environment, especially during her interrogation with the suspects.
Alfendi LaytonThe titular protagonist although he's more of a deuteragonist. A brilliant inspector of Scotland Yard and the son of Professor Hershel Layton. He has another 'side' to him where his personality changes into a more threatening and criminal-obsessed one.
- Anime Hair
- Beware the Nice Ones: Alfendi is nice and quite a mild-mannered gentleman like his father but every time he and Lucy confronts a true culprit in every cases, Alfendi will not hesitate to get rough, hotheaded and offensive on them but the most particular one in case 005, Mike De Bonair deliberately gets his last name wrong so many times. Alfendi now past his temper's limit threatens to cut his tongue out taking the former by surprise. Subverted however when it's discovered later on that his impulsive demeanor is another of his split personality closer to the original Alfendi.
- Fake Memories: "Placid" Layton claims that he is the one who killed Keelan while "Potty" Layton has no memories beyond the shooting. The fake memories are courtesy of Justin Lawson.
- Exposition Fairy: He essentially serves as a guide teaching you how to play the game.
- Good Cop/Bad Cop: Played With. He's both.
- Leeroy Jenkins: Averted. Alfendi is much more likely to think things over before jumping ahead, the only exception being case 9 where people's safety was at jeopardy if he didn't comply with Diane. However, the real Prof doesn't seem to be this, jumping ahead of his colleagues when Keelan Makepeace's location was first discovered, leading him to a trap .
- Jekyll & Hyde: Has another side which is more violent and even threatens to kill someone during interrogations.
- Mentor Archetype: To Lucy.
- Missing Mom: Which is the question of many fans.
- Nice Guy: Is generally level-headed and usually says things in the nicest way possible, despite some of the verbal insults some of the suspects give him. At least, until the other Alfendi comes out, much to Lucy's horror.
- The Professor: Serves as Lucy's mentor. Isn't too fond of the nick-name though.
- Tomato in the Mirror: His other rude personality is actually his original personality. The nice personality was a new personality that appeared after the incident where he was shot.
- Split Personality: Occasionally swaps into a more threatening personality, dubbed "Potty Prof". Turns out that this split personality is actually the original, with the current "Placid Prof" actually being created through brainwashing
- Super-Powered Evil Side: His other side often provides the key deduction that ultimately cracks the case due to it being able to think like a criminal.
The CommissionerThe... commissioner of Scotland Yard. His name is Barton. Yes, the same Barton who was a constable and worked under Inspector Chelmey.
- Adaptational Badass: Went from Chelmey's sidekick to commissioner of the Scotland Yard.
- Big Eater: In the ending credits, we see him eat a large plate of food.
- The Commissioner Gordon: Averted since the main characters are members of the police force. But played straight in the last scene of the game which is a Sequel Hook showing the Commisioner calling Amateur Sleuth Hershel Layton asking for help.
- Continuity Nod: He used to be Inspector Chelmey's sidekick.
- Cool Old Guy
Deputy Commissioner ChanThe Deputy Commissioner who talked with Lucy Baker in the prologue.
- Hypocrite: Somewhat of one when it comes to superstitions.
- Jerk with a Heart of Gold: Despite being incredibly discouraging to Lucy about working in the Mystery Room, he does worry about her safety. Mainly because the inspector she works under with has a split personality.
Florence SichA forensics analyst. She gets sick very often and as a result, goes around in an ambulance.
- Ill Girl: Is hooked up to an IV machine and sneezes quite a bit.
- Meaningful Name: Her surname is pronounced "sick".
- It also sounds very similar to "Forensic."
- Proper Tights With Skirt: Noticeable in her full profile picture.
Justin LawsonA burly Scotland Yard inspector and basically the polar opposite of Inspector Layton.
- Dirty Cop
- Hardboiled Detective
- Jerk with a Heart of Gold: Despite being responsible for at least half the Jigsaw Killings, he couldn't bring himself to kill Alfendi twice despite having the chance. He is also the one who suggested to listen to Alfendi's statements.
- Treacherous Advisor: While she does this to other people as well, Lucy generally confronts Alfendi's behaviour to Justin, who at one point said that if Alfendi mentions any specific people or names, that Lucy should secretly talk to Justin about it. Turns out he is the main reason why Alfendi has his split personality in the first place. When Diane Makepeace appears, Justin's paranoia comes out, as he's not sure what her father had said to her before his death. As he was already rattled with Lucy's reporting about Alfendi's behavior hinting the 'Real Alfendi' aka "Potty Prof" might spill more of the truth about that night, he couldn't chance Diane finishing the job and exposing him. So she had to be Killed to Uphold the Masquerade.
Dustin ScowersThe janitor who often picks up information about Scotland Yard during his duties.
'Sniffer' HagueA detective working under Justin Lawson. Apparently has a great sense of smell.
Hilda PertinaxAn Interpol agent working with Scotland Yard to find a murderer who stole an artifact.
- Dub Name Change: From Hilda Swanson.
- Lovable Alpha Bitch: Downplayed. She scolds Lucy for eavesdropping on her conversation with Alfendi despite it being quite loud, a bit bossy and was initially adamant of convicting Alfendi as the killer. However, she generally looks after Lucy, telling her Placid Prof was the fake persona and offering her comfort. By the end of case 9, she sides with Lucy on her theory about the true killer (though Hilda will begin to regret it if the player makes a mistake).
Sandy AldwichA woman found dead on her vacation at a five-star hotel. For some odd reason, her hand was found in her sandwich she ordered.
- Asshole Victim: She gets her money from her boyfriends and afterwards dumps them for another man.
- Dying Clue: Supposedly left one as she was apparently a fan of crime fiction. Too bad the very existence of the dying message gave it away as a fake.
- Gold Digger: Possibly. According to the killer, they used to be an item until he ran out of money, at which point she promptly dumped him. Also, her current lover is a wealthy businessman who is paying for everything on their luxurious vacation.
- Punny Name: Sandwich.
Jack PotsbyA loser who recently won a huge sum of money.
- Punny Name: Jackpot.
Gloria BlaiseThe lead actor of a play where she was the heroine.
- Driven to Suicide: She made it elaborate enough to frame Strapping for murder.
- Punny Name: Blaze of glory.
- Suicide by Cop
- Your Cheating Heart: Knew that Strapping was cheating on her with Knox. It distressed her far more than him, to the point that she plotted to commit suicide and ruin him.
Archie O'LogieA world-famous archaeology professor.
Dwight DreadA DJ who hosted a famous radio show.
- Asshole Victim: Once he found out de Bonair had been stealing profits from the radio station, he blackmailed him by having de Bonair give him money.
- Meaningful Name: Dread refers to his dreadlocks of his wig.
Anice BrewerA tea lady working at a radio station.
- Fangirl: Of Dwight Dread. Unfortunately, this was exploited to help the murderer kill the main victim and then pin the blame on her
- Proper Tights With Skirt
- Punny Name: A nice brewer.
- Red Herring: In-universe thanks to Sniffer.
- Spanner in the Works: Miraculously, she survives her poisoning long enough to leave a Dying Clue.
- Unwitting Pawn
Severino AgonniA mafia boss and head of the Agonni family.
- Asshole Victim
- Driven to Suicide: One possibility raised during the investigation. He wasn't a suicide.
- Properly Paranoid: He moved his gang's hideout to a warehouse in the docks because he feared that someone might kill him, and it turns out that he was right that someone wanted him dead. Specifically, Diane Makepeace, with the help from his right-hand man, Sassina, killed him because he betrayed her father at one point.
- Punny Name: Severe agony.
Barbarossa SassinaAgonni's right-hand man.
- Asshole Victim: Not according to Mistry's story of him.
- Dragon with an Agenda: Drugged and killed half of the members while he burned his boss.
- Punny Name: Barbarous assassin
Pearce 'Blood' BoyleThe Agonni family's strongman.
- Asshole Victim: A lot of people already wanted him dead because of his countless convictions for assault.
- Punny Name: Pierce boil
Angelo FassiA new recruit of the Agonni family and served as their pretty boy.
- Asshole Victim: For being a serial fraudster and larcenist.
- Bishounen: In-Universe, this is what his profile describes him as.
- Punny Name: Angel face.
Keelan MakepeaceThe serial killer responsible for the Jigsaw Puzzle Killings.
- Asshole Victim
- Bad Guys Do the Dirty Work: Some of his killings were requested by Justin when the Yard couldn't catch the criminals themselves.
- Kill the Creditor: Diane told Lucy that Keelan owed Pig a LOT of money, but one day Keelan flipped, and killed Pig rather than pay him back.
- The Killer Becomes the Killed
- Posthumous Character: Died from the incident four years ago
- Punny Name: Kill and make peace.
PigA millionaire who has a few associates with enough motive to kill him.
- Asshole Victim: At least that's what his colleagues think of him as.
Diane MakepeaceDetailed more on the Suspects section.
Bosco FelpsAldwich's lover and for business purposes, refused to tell the police his identity.
- Spell My Name with an "S": An in-universe example. Carrière thought his last name was "Phelps" instead of Felps".
- No Celebrities Were Harmed: This guy is practically Mr. T in a business suit. One of his arrows even says "I pity the fool!" and Lucy warned him to "B-A-round!" for further questioning.
Zach CarrièreA bellboy who brought Aldwich her sandwich.
- Fanboy: He is a self-admitted fan of crime fiction.
- French Jerk: Double Subverted. He tries to be helpful to Alfendi and Lucy until he learns that he is their main suspect.
- Great Escape: Seems to be discussing ways to escape prison with de Bonair in the credits.
- Starter Villain
Vera WipovskyThe hotel cleaner who found Aldwich's dead body.
- Red Herring: If you think that her description gives her the possible motive of killing Aldwich because her husband might have cheated on her with Aldwich, then think again.
- Team Mom: Has this relationship with her hotel employees.
- Your Cheating Heart: Worries her husband may be having her affair while she's away.
Goldie Potsby-MahnJack Potsby's wife.
- Black Widow: She had three husbands before Potsby and became rich from their life insurance. Despite that, she's apparently innocent according to the police and insurance companies. She messed up when it came to killing Potsby.
- Graceful Ladies Like Purple: Wears a slinky purple dress.
- Insurance Fraud: Suspected of such because her previous three husbands all died, and she got rich from the life insurance. She slipped up when killing Potsby, who had been spending money like crazy.
- Multi Coloured Hair: Her bangs are white, her side bangs are lavender and the rest of her hair is black.
- Pretty in Mink: Has a white fur coat◊.
- Shower Scene: There's a brief picture of her in the shower.
- Smug Smiler: Repeatedly gives this smile when discussing the murder weapon and how Alfendi and Lucy will never find it. And for good reason, since the murder weapon no longer exists.
Chase M. DownesThe muscular concierge of the apartment where Potsby lived in.
- Red Herring: His being muscular and joining an exclusive gym would make him match Goldie Potsby-Mahn's description of the burglar, but that's a dead end.
- Punny Name: Chase him down.
Buster NicksAn old serial burglar.
- Blatant Burglar
- Not Me This Time: He has a reputation as a petty criminal, and indeed was planning on breaking into a flat in Jack's building, but not his flat.
BingoPotsby-Mahn's new boyfriend. He's not actually a suspect, though.
- Beast Man: Looks less like a human and more like a lion standing on his hind legs. Goldie even keeps him on a leash like a pet cat!
- Meaningful Name: Named after the dog in the folk song "Bingo Was His Name-O".
- The Voiceless: Does speak sometimes, but it's basically barking.
Roscoe StrappingA famous actor partly known for his debauchery.
- All Men Are Perverts: He not only hits on Lucy but also assumes that Alfendi is getting some action from her. Because he believes that's what men do when they have cute female assistants.
- Demoted to Extra: In-Universe. In the credits he's shown as a lowly stagehand while Knox and Clegg are on stage. Presumably, the results from the case created a scandal that ruined his acting career or got him legally punished from hitting Clegg as part of his scheme to frame him.
- Drama Queen: Is really dramatic in his dialogue.
- Fake-Out Fade-Out: The conclusion seems to end once his heart meter breaks, and Strapping has no rebuttal to being accused of murder. But then the scene quickly transitions back to the interrogation room with his yelling encore, and he has a partially filled heart meter.
- Red Herring: He's set up as the murderer for much of the case, and though he did frame Clegg for the killing, he didn't murder anyone.
- Your Cheating Heart: Even though he's engaged to Blaise, he cheats on her with Knox, the supporting actress.
Destiny KnoxThe supporting actress for the play.
- Karma Houdini: By the looks of the credits, she doesn't get punished for lying about hearing Blaise scream.
- Punny Name: Destiny knocks (on your door).
Bray CleggA stagehand of the play.
- Ascended Fanboy: Went from stagehand to an actor as seen in the credits.
- Framing the Guilty Party: Strapping knocked him out to frame him, since he genuinely thought he switched the guns. He was wrong.
- Punny Name: Break a leg. Also possibly punning a kleig lighting on a stage.
Doug ScowersDustin Scowers's older brother and a student of O'Logie.
- Punny Name: Dig (and) scour. Another meaning of "scour" (apart from the one used for his brother) is to "seek" or "search", and he's an archaeologist.
Micah SasucasaA hotel owner.
- Bilingual Bonus: If you know Spanish, then try to say his name out loud.
Mariana EtistaA young woman working as a hotel employee. Actually a fake identity used by Diane Makepeace.
Chico CaretaA local of a village who volunteered to be a local guide for the archaeologists.
- Color Coded Emotions: He emotes through various colored masks. Red for Anger, Blue for Sadness/Nervousness, and Green for Joy/Laughter.
- Eloquent in My Native Tongue: Not only is his English broken and peppered with Spanish words, but he is also unable to read or write English as well. Justified in that he has only been learning English from Mariana for the past six months and is still learning.
- El Spanish "-o"
- =Karma Houdini: Apparently, he gets released, even though he killed O'Logie, because he was being manipulated by Mariana. In the credits, he's shown to be happily rolling a cart with a stone statue to the village, which surprises the villagers and Sasucasa.
- Love Martyr: He recognized Mariana's voice coming from the statue, which implies that he knew he was killing on her behalf, and not on that of a demon.
Dolly HollerdayA famous singer and a guest in Dread's show.
- The Ditz
- Giftedly Bad: She sees herself as a great singer, but her profile claims she's horrible.
- Ms. Fanservice: Her profile says she's able to distract from her bad singing by acting sexy onstage.
- Red Herring: Initially, not knowing much about the case, Alfendi supposes on a less-than-50%-chance she's the killer.
- Shipper on Deck: For Lucy and Alfendi, it seems.
Melody SmithA talent agent who became Hollerday's manager.
Mike de BonairThe friendly director of the radio show.
- Bitch in Sheep's Clothing: You see his true colours during the interrogation. He doesn't even know the name of the other co-worker he tricked and killed.
- Deadpan Snarker: During his interrogation, he will make snide comments about the Metro's efficiency.
- Great Escape: Seems to be discussing ways to escape prison with Carrière in the credits.
- I Never Said It Was Poison: He accidentally revealed that the container in which the poison was stored was a perfume bottle, when it had been shattered by Brewer after the murderer had been done with her.
- Ironic Echo: Repeatedly gets Alfendi's name wrong and then objects when Alfendi purposely messes up his name after he's exposed as the murderer.
- Meaningful Name: "de Bonair" equals "debonair".
- Multi Coloured Hair: Has two white bands of hair in the side.
- Smug Snake: Especially in the final part of his interrogation.
Randal MannThe sound technician of the radio show.
Uttar MistryThe delivery boy of a Dale Herl's Deli, a delicatessen.
- Blush Sticker: When he fanboys about the second-in-command mafia murderer and victim.
- Children Are Innocent: When Alfendi tells Lucy he is 100% certain Mistry is not the culprit, Lucy accuses him of thinking that because he's only a kid.
- Loony Fan: For someone who was worried about being killed by the mafia agent, he seems to be a big fan of him. He happily recounts Sassina's killing his fellow members in the credits.
- Punny Name: "Utter mystery". Not like you needed to ask.
- Spanner in the Works: If he hadn't been at the scene and inadvertently blocked one of Sassina's paths to escape, Diane would have had the opportunity to arrange the whole crime scene, since Sassina would have died anyway but not have locked himself up in the building.
Diane MakepeaceThe daughter of Keelan Makepeace.
- Affably Evil: Lampshaded by Lucy in Case 9.
- Avenging the Villain: Her acts against Layton are out of avenging her father's death
- Bait-and-Switch Boss: Despite being the main villain of cases 4, 6, 7, and 8 which build her up to be the final villain, she's killed offscreen at the start of the case 9 and confrontation against the final boss of the game is to uncover her killer.
- Cute and Psycho
- Daddy's Little Villain: Of Keelan Makepeace. The end credits show her and her father taking a picture of themselves robbing the vault of a rich victim Keelan just killed.
- Elegant Gothic Lolita: Played even more straight since she is an adult.
- The Killer Becomes the Killed: By the same way her father died no less. Justin couldn't tell what Keelan told her, so she was Killed to Uphold the Masquerade.
- Little Miss Badass: Lampshaded in her profile that despite her innocent appearance, she has international notoriety.
- Master of Disguise: She can disguise herself as Mariana with ease.
- Nice Hat: As part of her gothic lolita outfit.
- Perky Goth: Which generally adds to her creepiness.
- Punny Name: Die and make peace.
- Revenge Before Reason
- Serial Killer
- Soft-Spoken Sadist
- You Killed My Father: She wants revenge against Alfendi Layton for killing her father.
CatThe wife of Pig.
- Gold Digger: Although her profile states that she wants to claim the life insurance policy on Pig, she insists she isn't one as she wouldn't get the money Pig passed down to her. She's not the killer.
- The Unfought: It's entirely possible to play through the whole case and not interrogate her.
HenThe housekeeper for Pig's mansion. She found her master dead but decided not to report it and instead stole money from his wallet.
- But Thou Must!: Lucy will autonomously frame Hen when first contacting Diane, forcing the player to prove why she can't be the murderer.
- Red Herring: Her fingerprints on her wallet and her being increasingly annoyed by her master give her a rather plausible motive to kill, but the worst she is is a thief.
DogA trader who had borrowed money from Pig and so had to meet with Pig about his debt. His true identity is Keelan Makepeace, the infamous serial killer behind the Jigsaw Puzzle Killings.
- Hide the Evidence: The two pictures of him entering and leaving the room before Donkey arrives are the vital evidence to show that he had the opportunity to kill Pig. But in the actual investigation years ago, Keelan had those pictures hidden with help from Justin, so it was thus impossible to determine who the killer was.
DonkeyA shady jeweller who was supposed to meet with Pig about him knowing about the merchant's fraudulent practices.
- The Unfought: You never need to question Donkey, since he's innocent.