At a press conference held by Bruce Wayne to declare his opposition to Arkham City, Wayne is arrested by Hugo Strange's Tyger mercenaries and is himself imprisoned in Arkham City. Strange discloses his knowledge of Wayne's dual identity as Batman before releasing him into the criminal populace. While Strange prepares for the commencement of Protocol 10, Wayne obtains his crime-fighting equipment via airdrop by Alfred Pennyworth, allowing Wayne to become Batman. As Batman, he is first forced to save Catwoman from being executed by Two-Face who hopes to gain respect with her murder. After an assassination attempt on Catwoman by the Joker, Batman tracks the Clown Prince of Crime to his hideout in the Sionis Steelworks, believing he may know the truth behind Protocol 10.
Batman learns from Joker's doctor that the unstable properties of the Titan formula are mutating in Joker's blood, gradually eating away at his body and killing him. The Joker captures Batman and performs a blood transfusion on him; infecting him with the same fatal disease. Batman is forced to find a cure for himself and Joker, or they will both perish. Batman is informed that Mr. Freeze had been developing a cure but has since been kidnapped by the Penguin.
Batman pursues Penguin to his headquarters in the Cyrus Pinkney National History Institute: defeating his forces, his imprisoned monster Solomon Grundy, and ultimately the Penguin himself, before liberating Freeze. Freeze tells Batman that he has already developed the cure but that its instability renders it useless. Batman deduces that the restorative properties within the blood of Ra's al Ghul can complete the cure. Batman stalks one of Ra's assassins to his lair in Wonder City, leading Batman into a confrontation with Ra's and his daughter Talia — Batman's former lover. With Ra's blood, Freeze is able to develop a vial of working cure, but it is stolen by Harley Quinn before Batman can use it.
Batman returns to the Joker and finds him to be fully healed. While Batman and Joker fight, Protocol 10 is activated — A scheme to wipe out the entire population of Arkham City, destroying the criminal element of Gotham. The Tyger troops begin mass executions of prisoners, and Strange launches missile strikes from his base in Wonder Tower. A missile hits the steelworks, burying Batman under rubble. Before Joker can take advantage of the situation, Talia arrives and offers him immortality in exchange for sparing Batman's life. After he escapes with the help of Catwoman, Batman is convinced to put an end to Protocol 10 before pursuing Talia and Joker.
Batman infiltrates Wonder Tower and disables Protocol 10. Ra's is revealed to be the mastermind behind Arkham City, and mortally wounds Strange for failing to defeat Batman. With his dying breath, Strange activates Protocol 11 - Wonder Tower's self destruction. Batman and Ra's escape and Ra's commits suicide rather than risk capture. Joker contacts Batman, threatening to kill Talia unless Batman brings the cure to the Monarch Theater. Batman arrives as Talia escapes and kills the Joker. Talia offers Batman the cure, having intercepted it from Quinn, but she is killed by the real—and still dying—Joker. The second Joker is revealed to be Clayface, who had been masquerading as the healed Joker at the ailing villain's request.
Batman incapacitates Clayface, but Joker blows up the theater floor, sending Batman plummeting into Wonder City below. Batman destroys Ra's rejuvenating Lazarus Pit before the Joker can use it, and drinks a portion of the antidote. Batman debates curing his foe, but is attacked by the Joker, causing the antidote vial to smash. Batman maintains that in spite of everything Joker has done, he would have saved him. The Joker finally succumbs to his illness and dies. Batman carries Joker's body out of Arkham City into the winter dawn.
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