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Kuki Sanban, a reporter for Channel 6 Eyewitness News in Townsville City, investigates a crime wave by a group of criminals called the Moja Clan. At a dock at night, she sees the Mojas raiding cargo containers. After an unseen vigilante attacks the thieves, Kuki notices a symbol left behind. Kuki's supervisor Whoopi Thompson and her coworkers are oblivious to her story. Later while covering a charity event thrown by Sackz Industries, Kuki expresses gratitude to the company's CEO Derek Sackz, who was her late father's lab partner.

Frustrated by the vigilante, the Moja Clan's leader Mojo Jojo has the Moja Soldiers take hostages at a subway station in order to draw him out. Kuki, at the scene, becomes a hostage herself. Four mysterious figures arrive, take out the Clan, and free the hostages. Kuki follows them to a rooftop and is confronted by four superpowered teenage girls, they advise her not to tell anyone of them. As they leave, Kuki hears Buttercup and Blossom's names.

Kuki returns to her apartment and remembers "Project B", her father's science experiment, which involved four young female clones of Professor John Utonium named Blossom, Buttercup, Bunny and Bubbles Utonium. Unable to convince Whoopi of the girls' existence, Kuki is dismissed. Her coworker Otto Osworth drives her to Sackz's estate where she confides in him about her discovery. Sackz believes her and reveals that he, Professor Utonium and Kuki's father had been experimenting on a Chemical X created to cure disease, which was thought lost in the fire that killed her dad.

At the Professor's behest, the Powerpuff Girls bring Kuki to their home. The Professor explains Kuki had saved his superpowered clone daughters from the fire and freed them. The Chemical X gived the four of them develop superpowers. The Professor took on the role of their father, using Kuki's father as an example. After he proceeded to teach the girls in the fighting style. When Kuki reveals she told Sackz about her discovery of the girls, The Professor informs her that Sackz turned on her father and killed him.

Then, Mojo and the Moja Soldiers attack their home, defeating the Professor and incapacitating Buttercup while the other girls are captured. Kuki comes out of hiding and she and Buttercup plan to save the others. At Sackz's estate, he has the girls' blood drained in order to create an antidote to a deadly virus that Sackz hopes to flood Townsville with, believing he will become rich from people seeking his cure. Buttercup, Kuki and Otto storm the estate and free the other girls. The group then escapes the compound in pursuit of Sackz.

On a radio tower in the city, Sackz and Mojo plant a device that will flood the city with the virus while Sackz is preparing to convert the Chemical X to healing factor. Kuki and Otto subdue Sackz in the lab, while the girls battling Mojo on the roof. During the fight, the tower's support beams collapse. As the Powerpuff Girls try to keep it from falling and infecting the city, Kuki confronts Mojo with the chemicle x. In the struggle, the tower collapses and the Powerpuff Girls pull Kuki onto it with them, while Mojo falls to the street and is confronted by police. Believing they are about to die, the girls confess their secrets, while Buttercup gives an impassioned speech of her love for her sisters before they land harmlessly on the street. They vanish before the someone find them and return to the Utonium home, where they give the Professor the chemical X and he begins to recover.

Sometime later, Kuki meets with Otto, who tries and fails to ask her on a date. The Powerpuff Girls appear in a special modified "Powerpuff Van", and Bubbles accidentally blows up Otto's new car with a rocket. As police respond to the explosion, the Powerpuff Girls leave, but not before Bubbles tries to serenade with "Happy Together", much to her sisters' annoyance and Kuki's amusement.

Also see the Powerpuff Girls return.

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