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Gru, Lucy, Margo, Edith, and Agnes welcome a new member to the family, Gru Jr., who is intent on tormenting his dad. Gru faces a new nemesis in Maxime Le Mal and his girlfriend Valentina, and the family is forced to go on the run.

Gru and Lucy are overjoyed as they welcome their newest family member, Gru Jr. However, their happiness is short-lived when a new villain, the cunning Maxime Le Mal, emerges. With their family in danger, Gru and his team must utilize their wit and skills to outsmart the villain and protect their loved ones.—Oscar Serpa

Former super-villain and undercover Anti-Villain League (AVL) Agent Gru (Steve Carell) attends a class reunion at his former high school Lycée Pas Bon (which specializes in reaching Villainy to its students) where he reunites with Maxime Le Mal (Will Ferrell), a former classmate of his who has a feud with Gru for "stealing" his talent show act. It was the 9th grade, and Maxime had practiced for weeks. But Gru went before Maxime and performed the same dance number on stage. Although it is not clear if Gru stole the idea from Maxime or if they both had the same idea by chance.

Maxime is a French-accented, cockroach-themed super-villain who was an old high-school rival of Gru and seeks revenge against him and his family. Even at the reunion, Maxime makes fun of Gru, and he has no repartee to his witty comments. Maxime Le Mal also wins the Golden Alumni award, beating Gru.

During the ceremony, Maxime reveals he has enhanced himself with cockroach body parts, and Gru has him arrested by the AVL. Helped by his girlfriend Valentina (Sofía Vergara), Maxime escapes prison to get revenge on Gru and reveals his new invention, a gun that transforms people into cockroach hybrids, which he plans to use specifically on Gru's infant son Gru Jr (Tara Strong).

Days later, Silas Ramsbottom (Steve Coogan) the previously retired director of the Anti-Villain League, visits the Gru household to inform them that Maxime has escaped and that the AVL will have to relocate the Gru family to a safe house in another town called Mayflower under new identities. The family includes Lucy (Kristen Wiig), Margo (Miranda Cosgrove), Edith (Dana Gaier), Agnes (Madison Polan) and Gru Jr. Gru Jr is very close to Lucy, but does not share the same affection for Gru. The family is given only a few hours to pack everything and to be ready for relocation.

Meanwhile, the AVL takes in the Minions. Silas selects five of them to be augmented with superpowers under a new initiative called the Mega Minions. The five selected had been volunteered by the rest of the Minions group. However, the initiative is called off after the group unintentionally caused significant collateral damage in a nearby city.

In their new well-off residential town, Gru is assigned a safe house which is the most ordinary looking house of the entire town. Gru's new identity is of Chet Cunningham, a solar panel salesman and Lucy is Blanche, a hair stylist at an upmarket salon. Edith is Blair, Agnes is Britney, and Margo is Bree. Agnes is not comfortable lying about her identity to others, but Gru orders her to.

Silas says that Gru tends to stand out in a crowd and asks him to try and blend in for the sake of the safety of his family. On her first day at work, Lucy ends up burning the hair of the salon's most snobbish customer. The irate woman chases Lucy in the aisle of a shopping center and Lucy barely escapes.

Gru and his family meet their new neighbors, the Prescotts Perry (Stephen Colbert) Gru's arrogant neighbor who owns a line of automobile dealerships and Patsy (Chloe Fineman) Perry's Texan-accented wife, and their teenage daughter Poppy (Joey King).

Edith and Agnes attend a Karate Dojo, where the Sensei picks on Agnes as she is still not comfortable giving her name as Britney. As the Sensei yells at Agnes, Edith intervenes and hits the Sensei on his foot, breaking his little toe. All the other girls of the Dojo start respecting Edith now.

Maxime and Valentina search for Gru in their Cockroach shaped ship.

Poppy, an aspiring villain, recognizes Gru from his former career as a villain and threatens to blow his cover unless he helps her pull a heist to steal Lycée Pas Bon's honey badger mascot Lenny. Gru tries to reason that honey badgers are vicious creatures who eat cobras and bees for breakfast. But he has no choice as otherwise Poppy would expose Gru's real identity to the neighborhood.

With the help of two Minions and Gru Jr., they successfully steal Lenny from the school. There are several obstacles as the Minions forget Gru's gadget bag in the hot air balloon, Gru Jr continues to pester Gru, and Poppy is wide eyed at all the portraits of all the villains who passed out from the school. Gru Jr releases the badger, who was hung in a cage in the principal's office, and it ends up attacking Gru. Poppy accidentally activates the alarm, which wakes up principal Ubelschlecht. Gru Jr tames the badger by feeding it his own mashed food supplies. The team manages to escape the school in a flying machine, after avoiding being trapped by Ubelschlecht. However, the team is unaware that Lenny's collar contains a tracker. Ubelschlecht (Chris Renaud) identifies Gru via security camera footage and contacts Maxime to inform him about Gru's whereabouts using Lenny's collar.

The next day, while Gru and Lucy are away at tennis practice with Perry and his wife Patsy, leaving the girls and Poppy to watch over Gru Jr., Ubelschlecht visits the safe house to interrogate them about Gru, which makes Poppy promptly leave carrying Lenny. Gru and Lucy then rush back home after being alerted by Margo and they all fight Ubelschlecht. Lucy also signals the AVL who calls the Mega Minions out of retirement to stop Maxime.

In the havoc, however, Gru Jr. leaves the house and is spotted by Maxime and Valentina, who kidnap him. Lucy and the girls stay behind to handle Ubelschlecht as Gru decides to save his son with help from Poppy.

Using a flying car, Gru and Poppy chase after the ship, with Poppy getting Gru onto the ship. Gru hijacks the ship, causing it to crash land onto a building under construction. Maxime reveals he has brainwashed and transformed Gru Jr. into a human-cockroach hybrid and they fight Gru off. Gru is wounded, but his encouraging words to his son manage to break Gru Jr. free from his brainwashing state and he attacks Maxime instead. Gru punches Maxime, and he falls off the building to the ground, where he is further squashed by the Mega Minions and a whole army of animals, who arrive in turn. The AVL then arrest Maxime and Valentina.

Gru Jr finally warms up to Gru and calls him Dada.

Reunited with Lucy and the girls, the family return to their original home, where Dr. Nefario (Romesh Ranganathan) reverses Gru Jr.'s transformation. Sometime later, Gru visits Maxime in prison to talk and settle their differences. The former rivals then put on a performance of "Everybody Wants to Rule the World" at the AVL prison for the prisoners, attended by villains from the before, while Poppy is accepted at Lycée Pas Bon.