Number 5's repeatedly shocked, horrified reactions to several tests and treatments, especially the horror movie test ("OH LORD! OH LORD! OH GOD! WHY?!") and "extreme solitary." The poor man seemed to find everything that VAL did to be incredibly disturbing, yet still made it all the way to the final treatment.
Number 4 misplacing his Companion Cube ball because he had imaginary friends to play chess with.
During the rubber-ball-counting test: "Guests, I hope I do not distract you by talking to myself." The comedic timing of Number 3's immediate reaction just makes it.
Number 3 being scared of the slot in the Pod.
Number 3, given exemption from a Treatment, takes time to go completely crazy.
VAL: Hmm. That is a side of Number 3 I have not seen before.
Number 5 complaining to VAL about forming a union with Numbers 3 and 7 after ranting about how cold it is.
VAL asks the guests to draw self-portraits of themselves as they were when they entered, then as they see themselves now. Number 5 draws stick figures of himself under a happy sun, then one of him freezing.
Number 5: I was bald then and warm!
[on the 'after' picture]
Number 5: Happy and enlightened but freezing!
Number 5 in general is a crowning person of funny, from his reactions to everything to his over-the-top facial expressions.
In the finale, Number 5 comments that when people say you should be strong and ready for whatever the world sends your way, the world does not usually send a box of spikes.
4 of the guests stayed in the box for over 4 hours, far past the 3-hour stipulation.
The guests were kept without food or sleep for 30 hours, and some became... unhinged.
Number 8: I am hungry~ I am gonna eat my arm~
Number 5's memetic "Give me. The pizza. Now." Despite the line never appearing in the show outside the intro, it became so famous that he was asked to repeat it at the reunion.
VAL offered the guests paints to draw on the walls of the Pod; they reacted with delight. Then it cut to Number 5.
Number 5: [gloomy] If only I knew how to paint.
Number 7 reacting poorly to the brick/speech test.
During the ball gag treatment, Number 8 attempted to tell VAL "You son of a bitch!" in a muffled voice, to which VAL responded, "You want to give me a kiss?" He also declared, "This sucks!" to which VAL said in a cheery voice, "I agree . . . this rocks!"
Number 8's suffering in that treatment is comedically sociopathic. A particularly notable instance is when VAL allows him to retrieve a water bottle. "Now what?" "You tell me."
The guests cheering for their pet mice in episode 5 is heartwarming...except when the mice fall asleep or go the wrong way.
The guests taking out their frustrations on the much-reviled meal bars in Episode 5, including yelling at it, stomping on it, and throwing it into walls. Number 9 even gives it a Drill Sergeant Nasty rant.
On the Reunion Show, Number 1's nephew asked him why he passed gas on TV.
Number 9 whistling as he casually does laps on a ring of rocks.
When Number 6, who hated technology, leaves, VAL wishes him well...then says "Score one for technology."
Number 8 trying to sit on a spiky lawn aerator during the stool challenge.
VAL allowed the guests to make personal videos of themselves with a video camera. Afterwards, she then told each of them that their video diaries showed strong evidence that they were going insane. The only one who fell for it was Number 3, who became honestly worried that VAL was telling the truth. Shortly after, VAL revealed the truth. "This was not a test of sanity, but a test of gullibility."
Val in general gets a personality upgrade that seems to make her a lot more suggestive and willing to "come down" to the players' levels, even cursing at them in a few cases. Some of the better high points:
Number 3's Companion Cube robot dog doesn't work, prompting her to shove it back into the meal slot.
Numbers 4 and 7's increasingly comedic one-liners in the finale, punctuated by an "Oh, snap." from VAL.
Number 8 adding a bunch of AKAs to his name at the start, all of which VAL said were incorrect.
Number 8:...AKA Number 8.
VAL: AKA correct.
The guests engaged in a treatment in which they had to sweat as much as possible and then collect the sweat, the winner being the one who collected the most. Most of the guests managed 2-6 vials of sweat; the winner, Number 8, got 25. When the rest of the guests heard about the ridiculous amount Number 8 got, their shocked expressions were hilarious. "25?! Did they pee in the thing?!"
Number 7 mentioned she hadn't sweated that much since she was locked in the trunk of a car. Cue Record Needle Scratch and VAL's monitor reading TMI.
In the bartending Treatment, Number 1 wished for anything but pickle relish and refried beans. Cue another guest mixing exactly that.
One of the guests requesting a cigarette, and VAL complying by giving them one . . . without a lighter.
Number 6 won a challenge and got a bowl of peanuts and two beers as a prize. VAL then allowed the other guests the chance to beg Number 6 to share, with VAL as the go-between. However, VAL intentionally misinterpreted their messages, so Number 2's request of "Will you please share your peanuts with me?" became, "Will you please share your penis with me?" After he realized the misunderstanding, Number 2 tried to correct it, specifying that he'd said peanuts. VAL then reported the correction to Number 6. "Number 2 actually wants your nuts."
The guests were asked to hit a punching bag a certain number of times and report their score, with the one who reported the highest score getting a hot chicken dinner. Number 2 made up the ludicrously impossible number of 50,999,999,999 hits, and won the challenge. However, when he went to claim his prize, he found a tasteless vitamin block instead of a chicken dinner, and exploded with outrage, demanding to know where the prize he was promised was. VAL then responded, "You lied during my challenge, so I thought I would lie to you as well." Meanwhile, everyone but him got the dinner he was promised.
Number 5 going crazy in the Vallors auction to the point where he thinks none of the other contestants are real, nearly hits the red button for no reason, and buys the same item twice.
Number 2 is so demoralized after learning he wasn't the first to escape from Valcatraz that he goes back into his cell.
Number 2: ...I'm going back in here...Life was so much simpler inside the jail.
In the same treatment, Number 2 spends a lot of time trying to get a bag that he believes will hold a key to his cell. When he makes it...
Number 2: ...Cookies!? Are you freakin' kidding me!?
Number 7 losing the rope outside her cell and having to stretch with her foot and the bag of cookies to reach it. She loses a shoe in the process.