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The very first scene in which we see Gru. Wearing a grim expression on his face, he's assembling a gun. He points it, pulls the trigger... and out comes a balloon shaped like a unicorn. Then the viewpoint pulls back and we see that he's preparing Agnes' birthday party.
"I'm sorry, I did not see you there."
"You know, you really should announce your weaponsafter you fire them, Mr. Gru. For example... *shocks Gru*Lipstick taser!"
The minions making fun of Silas' name.
Silas: I am the league's director, Silas Ramsbottom.
Lucy unleashing a flurry of martial-arts moves... on falling cupcakes. "Wasn't expecting that... or was I?" She then boasts her moves are "a mix of jujitsu, Krav Maga, Aztec warfare, and kruuumppppiinnnn!"
Margo's expression when she saw Antonio for the first time.
Especially when she tries to chat with him.
Antonio: Cool glasses.
Margo: Uh... [chuckles nervously]
Antonio: Iím Antonio.
Antonio: I was just going to get a cookie. Care to join me?
Margo: [nervous] Uh,... sure. Iím... Margo.
Gru: Kyle, NO! Do not do your business in the petunias! (picks him up and drops him in the neighbor's yard) There. Those are Fred's. Go crazy.
''Kyle's pee incinerates Fred's flowers.
Gru: (Wicked chuckle) Good boy....
The minions holding auditions for their upcoming movie, but they're continuously blocked by a black wall.
At the end of the audition, in the midst of trying to swat a moth, a minion begins to fall and another minion tries to catch him but falls with him. The last minion shrugs like, "Screw this," then jumps after them, screaming, "BAILAAAAAA!"
Gru and the minions send Dr. Nefario off with a "21 fart gun salute". This leads to:
Dr. Nefario: I counted 22.
(He and Gru look at a minion who laughs bashfully)
And also, the way Dr Nefario exits Gru's evil lair. His scooter-jetbike is still very...slow.
Dr. Nefario: This may take a while.
Agnes' excitement for the arrival of the fairy princess.
As well as her absolute glee when she shows up.
Gru trying to find the serum in Eaglesan's shop, using a device Lucy has planted on him. He has to walk around the shop, get near everything he can, to find it... problem is it's on his lower-abdomen so when he tries to pick up a signal he's pelvic-thrusting at everything.
Basically anything to do with the minions. From the 'Brave Knights' scene to the 'Minion Island' scene to the 'Ice Cream Party' scene to the 'Hunter Posing with Dr. Nefario and El Macho' scene. I swear, the developers probably spent more meetings trying to pick the best minion scenes to put in the film. So much so that they're gonna make minions film. Which would probably be about why the minions end up with Gru and why they adore him so much.
The robotic way that Agnes recited her poem. Made even funnier by Gru's lampshade of it:
"Wow. That—was...something else! I really liked the way you smiled at the end—? Let's try this one more time, but a teensy bit less...like—a zombie, o-kay...?"
The Minion Golfing scene. Totally a throwaway gag, but hilarious. Minions A and B put the girls to sleep. Minion A says something to Minion B, and then the movie cuts to Minion A dresses in a golf outfit, with Minion B laying on his back with a golf tee/ball in his mouth. Minion A takes a couple practice swings at the ball, looks directly at the camera, and then lowers the club to the level of Minion B's head. He winds up...and is interrupted.
The minion with the golf club is Kevin (who attacks the house later on in the film once he's mutated) while the other one (the shorter and chubbier one) is Jerry. You can hear Gru say their names in the 'Ice Cream Party/Irish Drinking Song' scene.
Gru's attempt to do the dance-step password for El Macho's underground lair, only to keep tripping up and triggering booby traps.
The chip-and-guacamole hat.
Anything to do with El Macho's pet attack chicken.
Gru: I really hate that chicken. (The chicken activates The Rocket with Gru and Lucy on it )
When Gru uses Lucy's lipstick taser to take down the mutated Eduardo.
Gru: Lipstick taserrrr!
Lucy: Aw! He copied me.
The conversation when Agnes sees Edith grabbing coins from a wishing fountain.
A quick example: when the AVL is showing Gru the footage of the serum's testing, he sees the bunny mutate into a horrible monster. The Minions accompanying him look horrified, but if you look at Gru, he has this big smile on his face like it's the coolest thing ever.
However, when the monster bunny begins to attack the scientists, Gru's face is priceless.
Gru: You don't usually see that in bunnies.
Lucy's Love Epiphany on the plane. After seeing Gru in both magazine ads and in her fellow passengers, she decides to just open the plane's window and jump out with a hang-glider. But before that she turns to the stewardess she was talking to and cheerfully shouts "Thank you, Gru-Stewardess!"
Stewardess: *confused* You're welcome!
The cat being sent back down to earth. It was like "We have no use for cats here. Go forth and get adopted."
The minions getting abducted by the ice-cream truck. Especially the last minion who tried to run for it but was distracted by an ice-cream and then got taken too.
Oh, and the funny way the minions were running past the young children towards the ice-cream truck.
"GELATO!!!" Which is Italian for Ice Cream.
Gru simply plowing into every other car in the lot at the Cinco de Mayo party.
The entire date scene, including the part where they carry Shannon home.
Lucy: Looks like your date's been shot with a mild moose tranquilizer. I'm winking at you 'cause that's what actually happened.
Blink-and-you'll-miss-it example. Gru and Lucy sneak into Eduardo's restaurant after closing hours to see if they can find the serum. They find something, but it turns out to be a jar of "secret salsa." Cut to Eduardo returning...and then cut back to Gru and Lucy, who apparently figured they might as well have a snack, because they're dunking chips in the salsa and eating it instead of continuing the mission.
"HE'S SO FLUUUFFFYYYY!!!" (Sadly, the quote was only in the junior novel.)
"I'M SO HAAAPPPPPPYYYY!!!"
The scene where Gru is rowing and one of the minions directs him through a megaphone.
Which pops up again later, and is probably funnier because it's a Minion directing another Minion to pick up Gru and Lucy...and rowing right past them.
Gru: "They'll be back."
"Uh, hi everybody. I'd like to make some toast."
The entire "never grow up" scene:
Gru:(Kisses Edith's forehead and playfully bops her nose) Good night, Edith. (Walks to Margo and kisses her forehead) Good night, Margooo-woah-woah-woah-woah, hold the horses, who are you texting?
Margo: No one, just my friend Avery.
Gru: Avery... eh, Avery? Is that a girl's name or a boy's name?
Eduardo's mountain top manor is quite ridiculous from an architectural standpoint. It's a sprawling complex designed for a much bigger site; a good chunk of the estate is suspended off of the flattened peak by wooden beams.
Also the previous scene when a minion was tasting the jelly, then realizing how awful it tastes, he screams and smashes the jar on to floor and all minions begin to run amuck.
The entire introduction flashback for El Macho is Testosterone Poisoning-ridden HILARITY, but especially clever is a subtle Funny Background Event within it that involves a fellow taking a siesta...who doesn't even stir when El Macho punches out the door right near where the man is sleeping!!!