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A missile hits the steelmill, burying Batman under rubble. The TYGER troops begin executing inmates as Strange launches missile strikes on Arkham's denizens from his base in Wonder Tower. While the two fight, Strange activates Protocol 10, which is revealed to be a scheme to wipe out the entire population of Arkham City and destroy the criminal element of Gotham. When he returns to the Joker, Batman finds him restored to health.

With Ra's al Ghul's blood, Freeze is able to develop an antidote, but it is stolen by Harley Quinn before Batman can use it. Batman tracks one of Ra's al Ghul's assassins to his underground lair, leading Batman into a confrontation with Ra's and his daughter Talia, Batman's former lover. Batman deduces that the restorative properties of Ra's al Ghul's blood can complete the cure. Freeze tells Batman that he has already developed the cure, but its instability renders it useless.

Tracking Penguin to the Cyrus Pinkney National History Institute, Batman defeats his forces, his imprisoned monster Solomon Grundy, and ultimately the Penguin himself, before liberating Mr. Freeze, who had been developing a cure but has since been kidnapped by the Penguin. Desperate to save himself and innocent citizens, Batman seeks out Mr. Joker also reveals that Gotham hospitals have been poisoned with his infected blood. Joker captures Batman and performs a blood transfusion on him, infecting him with the same fatal disease. There, Batman learns that the unstable properties of the Titan formula are mutating in Joker's blood, gradually killing him. After Joker attempts to assassinate Catwoman, Batman tracks him to his hideout in the Sionis Steelmill, believing Joker may know the truth behind Protocol 10. He first saves Catwoman from being executed by Two-Face, who hopes to gain respect by murdering her. While Strange prepares to commence "Protocol 10", Wayne obtains his equipment via airdrop from Alfred Pennyworth, allowing him to become Batman. Hugo Strange discloses his knowledge of Wayne's dual identity as Batman before releasing him into the prison's criminal populace. At a press conference held by Bruce Wayne to declare his opposition to Arkham City, TYGER mercenaries arrest and imprison him in Arkham City.