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Apollo Justice is a rookie attorney with only four cases under his belt. His mentor was Kristoph Gavin, a skilled but mentally unstable defense lawyer whose pride and paranoia led him to commit multiple murders. He is now the protégé of legendary attorney Phoenix Wright. Apollo, who recently discovered that he is the son of Thalassa Gramarye, former member of a troupe of magicians, and the half-brother of Trucy Wright, Phoenix's adoptive daughter, possesses a bracelet which allows him to detect when people lie.
- Brought Down to Normal: In this trial, he can't use Perceive on people on account of losing his bracelet.
- Dude, Where's My Respect?: Like Phoenix, hardly gets any respect.
- Good Cannot Comprehend Evil: He gets a little one-to-one talk with the Big Bad towards the end, and makes it clear he can't even come close to understanding said Big Bad's "deranged mind".
- "The Reason You Suck" Speech: Gives one to the Big Bad.
- Was It All a Lie?: He's about as shocked as everyone when he realises that Rhea's the killer. When Rhea taunts him about how she'll get away with her crimes, he switches from her best pawn, to her greatest enemy.
Ema Skye is a detective from the Los Angeles Police Department leading the initial investigation in the murder of Robert Erlenmeyer. After the death of her parents, she was raised by her elder sister, ex-Chief Prosecutor Lana Skye. She dreamed of becoming a forensic scientist, though she did not succeed in qualifying for the position, and was instead hired as a detective.
- Big Damn Heroes: Arrives in the courtroom just before the verdict is entered with three crucial pieces of evidence that help corner Rhea.
Phoenix Wright is a legendary ex-defense attorney known for his nearly spotless record and tendency to turn hopeless cases around. His career went off after the murder of his mentor, Mia Fey, and he quickly rose to fame over the next few years. Unfortunately, it was all cut off abruptly just a few months after the trial of Iris of Hazakura Temple, when he was caught with forged evidence in the trial of Zak Gramarye. After clearing his name and catching the culprit behind his disbarment and several murders, Kristoph Gavin, he has left for Borginia with his adoptive daughter Trucy and her mother, Thalassa Gramarye, leaving his protégé Apollo at the office.
- Demoted to Extra: Due to being away nearly the entire time, he's mostly delegated to this role, occasionally contacting Apollo by cellphone.
- Put on a Bus: He's traveling in Borginia until the ending.
The Honorable Judge Chambers, better known simply as "the Judge" or "Udgey". One of the city's most famous judges, he now finds himself on trial for the murder of Robert Erlenmeyer, the Mysterious Bust Killer in revenge for the death of his brother Arthur.
He gets acquitted, but is murdered later that night after witnessing Rhea trying to kill Erlenmeyer.
- He Knows Too Much: The reason he was killed was because he saw Rhea about to kill Erlenmeyer.
- His Name Really Is "Barkeep": His first name is "Judge".
- Intergenerational Friendship: With Rhea, whom he met while on the Mysterious Bust Killer case. At least, that's what she wanted people to think.
- Killed Off for Real: He is murdered by Rhea the very night after his acquittal.
- Spanner in the Works: To Rhea Wits. He accidentally takes sleeping medicine instead of tranquilizers, then heads to the cemetery late at night. He gets killed, but this gradually brings about the killer's downfall.
- Unwitting Pawn: To Paul Strings; his visit furthered the plan to spring Erlenmeyer.
- Villainy Discretion Shot: In a sense, is applied retroactively. Chief Justice Strings argues that the way he handles cases may well have gotten innocent people convicted.
Better known as "the Judge's brother", Arthur Chambers was the Canadian judge who presided over Mia Fey and Miles Edgeworth's first trial. He was a victim of the Mysterious Bust Killer.
- Bus Crash: He was killed between his appearance in the official games and this case.
- Posthumous Character: Has been dead for some time before the beginning of the case.
A former police officer, court bailiff and Blue Badger mascot, Mike Meekins is a guard at the Clooney Prison Asylum who was on duty when Judge Chambers visited Robert Erlenmeyer.
- Too Dumb to Live: In traditional fashion, as he let Chambers past.
Jake Marshall is a former detective of the Los Angeles Police Department. Demoted to patrolman after the SL-9 case, he turned himself in two years later for stealing evidence and assaulting Officer Mike Meekins and impersonating the late Detective Bruce Goodman in an attempt to reopen the case. He is now a deputy of the Los Angeles Police Department, and shows up to testify in the second day of the second trial in place of Detective Skye, who is out gathering evidence.
Rhea Wits is a bright and cheerful young woman who happens to work in the death industry. Her brother Nathan was brutally murdered by an infamous serial murderer, the "Mysterious Bust Killer", and she had to testify in the trial of the alleged culprit, Robert Erlenmeyer, where she befriended Judge Chambers who like her had lost of a member of his family to the murders. When Judge is arrested for the murder of Erlenmeyer, she decides to help Apollo investigate the case, convinced that there is more to the story than meets the eye. She is actually the real Mysterious Bust Killer, and the true culprit of the case.
- Bare Your Midriff: Wears a red top that shows off her shoulders and midriff.
- Beneath the Mask: What makes her so repulsive, and yet at the same time, unconsidered as a suspect is the sheer lack of any reasonable motive for the crimes. A biology teacher snapping from reliving a childhood trauma is leagues easier to swallow than a young, seemingly well-adjusted woman deciding to kill several people because she believed they weren't really alive. Paul Strings unintentionally compares her to the Joker with the Burmese Bandit quote, and all things considered, its fairly fitting.
- Bitch in Sheep's Clothing: Basically, she seems nice enough when you first meet her, but it turns out, there's not a nice bone in her body.
- Breaking Speech: Gives Apollo one to eat up his time during the recess. It also serves as a distraction to keep him from devising arguments against her.
- Darker and Edgier: She manages to surpass every other canon Big Bad before her. As horrible as they were, Matt, Dahlia and Kristoph had human, if petty excuses for their crimes, and each personally killed at most three to four people. Rhea outdoes all of them by being a Serial Killer of twelve people for no other reason than the thrill of the crime, and to boost her ego.
- Evil Cannot Comprehend Good: As pointed out by Apollo, she's clearly incapable of loving anything or anyone, even herself.
- Expy: Being a Plucky Girl and The Watson, nevertheless involved in a business regarding the dead, she seems to be channeling Maya Fey. In reality, being a Bitch in Sheep's Clothing and the Big Bad, her tendency to refer to her 'love interest' by a pet name, and the stinger's implication of her developing a vendetta against the defense attorney who exposed her makes her a clear cut Expy of Dahlia Hawethorne. Her physical appearance is even based off of the concept art for Dalhia Hawethorne◊.
- False Friend: With Judge, whose brother she murdered, and with Apollo, who she's stringing along for her own ends.
- Flower in Her Hair: She wears a single red flower in her hair. She still has it while she's in prison, though it's been shredded to pieces.
- Freudian Excuse: Defied. Nothing in the story suggests she suffered any personal tragedies beyond losing her brother that would encourage her to become a killer, especially when you consider she killed him in the first place. When she finally shows her true colours, one gets the impression that one day, she just thought to herself how fun it would be to kill people, and decided to give it the old college try.
- Insane Troll Logic: She thinks murder is the highest form of living so logically, it should make her a would-be higher life-form. And why is it okay to kill people? Because they're apparently "not really living."
- Intergenerational Friendship: With Judge Chambers. Or so she wants everyone to think.
- It's Personal: With Robert Erlenmeyer for killing her brother. Subverted when it's revealed that she was the one who murdered her own brother. The Stinger implies that she has developed this relationship with Apollo for exposing her.
- My Card: Gives Apollo her business card when they first meet.
- Plucky Girl: She's somewhere between this and a Shorttank for most of the game. Things kind of go downhill after The Reveal.
- Significant Anagram: Rhea Wits = It Was Her.
- The Sociopath: To put it mildly. Hoo boy, she'd give Dahlia, Matt or Kristoff a run for their money! She even killed her own brother just to throw the police off, and get involved in the case, and as Apollo points out, she's clearly incapable of loving anything or anyone, even herself.
- Straw Nihilist: Her reason for killing people is essentially that she thinks killing is the highest form of living — so logically, killing as many people as she likes is not only perfectly okay, but makes her a would-be higher life-form. And why is okay to kill those people? Because they're apparently "not really living".
- Time-Limit Boss: Manages to convince Gerald Strings to put a time limit on the final part of the trial. After all, the witnesses can't take all their time coming up with what to say, so it's only fair that the defense would have a time limit.
- Villainous Breakdown: She grabs a shovel from hammerspace and digs a hole in the courtroom floor.
- Walking Spoiler: There is a vast abyss between who Rhea seems to be and who she truly is. If you knew more, it would be massive spoilers.
- The Walrus Was Paul: As it turns out, there was no motivation at all behind the sheet music and fake naked photos of the victims arranged in the shape of a bust. She just put these things in her crime scenes to mess with the police.
- What the Hell, Hero?: When Apollo decides to defend Erlenmeyer, she's quite outraged. But it turns out that it's mainly because she wants him to be convicted for her crimes.
Robert Erlenmeyer is the victim in this case, and is also known as the Mysterious Bust Killer. Prior to his killing spree, he was once a biology teacher. He's not actually dead, he was broken out of prison by his brother. Furthermore, he's not the real Mysterious Bust Killer, and he's accused of murdering Judge Chambers the day after the latter's acquittal.
- Affably Evil: His guard admits that he doesn't cause any trouble, and his psychological profile reveals that he was well-liked by his neighbours and students before confessing to being a Serial Killer. Subverted. Even though he is insane he is a genuine Nice Guy, and is really a patsy for the real killer.
- Asshole Victim: Serial killer killed to avenge a victim. Subverted on both counts — he's still very much alive and possibly only killed a cat by accident. He's mentally ill, not murderous.
- Freudian Excuse: The death of his cat, which he believed to be his fault, drove him into madness. But it's subverted, as even mad, he never killed anyone
- Nap-Inducing Speak: Shawn O'Fisher mentions that he had a hard time staying awake while listening to Robert's dull and monotonous voice. Apollo later uses this as an explanation as to how the Judge was made unconscious without any blunt trauma or drugs.
- Never Found the Body: Because he's not actually dead.
- Posthumous Character: Subverted. He's actually alive.
- Word-Salad Humor: How he speaks, due to his insanity. The "humor" part is averted for the most part: the game does a very effective job at playing it for drama. Additionally, some of his gibberish actually makes sense once you know the truth. For example, when he says that "The strawberry killed the plum", he's referring to Rhea, who wears red clothes.
Shawn O. Fisher / Bob Erlenmeyer
Shawn O. Fisher is a guard at the Clooney Prison Asylum who is Meekins' superior. He is also Robert Erlenmeyer's twin brother Bob, and he collaborated with Paul Strings to break him out of prison.
- Disc-One Final Boss: He's set up as the culprit for killing Erlenmeyer, but it turns out the killing was faked.
- Jerk with a Heart of Gold: Despite his abrasive personality, he deeply cares for his brother.
Paul Strings is the Chief Justice of his State's Supreme Courtnote , and quite a towering presence. A self-taught man who comes from a poor family in the countryside with an abusive father, he is an avid reader who is fond of quoting famous philosophers and intellectuals, and has a love for everything ancient. Even if he is not in charge of the Judge Chambers case, he seems to be strangely involved with its events, and appears to have a history with the defendant. His son, Gerald, is Judge's replacement in court. He was also the mastermind behind the plan to break Erlenmeyer out of prison.
- To Be Lawful or Good: He's faced with this regarding Erlenmeyer. He chooses to break the law by freeing him, but accepts his punishment once he's caught.
A tough veteran prosecutor who's seen it all, yet never lost her drive. She and Judge used to be lovers in university, but she left him to concentrate solely on her career. With her unflinching determination to fight corruption and hard-nosed methods, she made a name for herself, but also dangerous enemies. As a result, she suffered a depression and had to start a new life in the Virgin Islands, where she founded a family. However, the mafia found her there, and killed her husband and kids. She returned to her country with a passion, and when she learnt that her former fiancé's brother had been murdered, she vowed to herself to do anything in her power to bring the Mysterious Bust Killer to justice to redeem herself.
- Amoral Attorney: Unlike Edgeworth, Manfred and Franziska, this is not evident at first glance. It comes up when Poole admits that she fabricated a psychiatrist's report to get Erlenmeyer the death penalty.
- But Now I Must Go: Is willing to accept the consequences of her forging the report, although she stays around for another day after the Judge is killed.
- Crusading Widow: Her husband and children were killed by the mafia, leaving her with a passion to bring justice to criminals.
- First-Name Basis: After Judge's trial ends, she allows Apollo to call her by her first name, but then retracts that during the second investigation phase.
- Troll: Messes with Gerald Strings when he arrives late due to walking in the wrong courtroom.
The son of Chief Justice Paul Strings, he is the replacement judge in charge of Judge Chambers' trial. He is relatively new to the courts.
- Cloudcuckoolander: Firmly established when he goes into an empty courtroom by mistake.
- Everyone Has Standards: A Cloud Cuckoo Lander, and something of a careerist, who's willing to indulge in unorthodox court procedures to improve his reputation. Then Rhea reveals what she really is and his only reaction is sheer, unadulterated disgust and an immediate Not Guilty verdict for the defendant. After the trial, he decides to retire as a judge partially because he realised that he unintentionally aided a Serial Killer out of his attraction to her, and to further his career.
- This Is Unforgivable!: He slips into this after the Big Bad is exposed.Gerald Strings: Get that... that... thing out of my courtroom, this instant! (...) BAILIFF, I SAID THIS INSTANT!
Robert Erlenmeyer's defense attorney.
- Insistent Terminology: Takes issue with the Mysterious Bust Killer's gender. It turns out that the real killer is a woman.