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One of the easter eggs involves looking into murky water for at least forty seconds. Upon doing this, a massive octopus-like creature swims by, glowing orange eyes fixing on you for a second. Try it again, and it attacks you with a tentacle, just barely missing its mark.
During one of the glyph puzzles, the player drags a fire-like icon over an image. They are then (likely) caught off-guard when the ambient silence maintained through most of the puzzle sections is abruptly interrupted by the sound of agonized screaming, with an image of Joan of Arc burning at the stake, with the words THEY BURNED JOAN ALIVE over it. This disappears quickly to move onto the next part of the puzzle, but when you're playing all on your own in a dark basement with the volume up, it really screws with your head...
Hell, the glyph puzzles in general!
Especially the one when you hear Subject 16 talking about killing and fucking people and completely losing his mind, much more so then the previous entries.
The Martyrs puzzle when you're going through the puzzle, the sound it makes isn't a beep, but a heart beating faster and faster as you get closer to the Piece of Eden, and when you ID it - flatline.
That poor fucking elephant. It's not fake. Thomas Edison really did that. Just to smear Nikola Tesla. IRL, there's no proof that he was a Templar, but there's the proof he was a dick.
The Poison Blade. It's victims begin feeling uneasy at first, but before long, they start thrashing on the ground and begging for help. No wonder Altaïr called poison a coward's tool!
Although they're practically part of the background, there is something particularly chilling in the outfits of the Plague Doctors
The "acquittal" of Giovanni, Federico and Petruccio Auditore... because it's one thing to see already-dead hanging corpses in a video game, it's a completely whole 'nother to witness the execution without a discretion shot, except for Petruccio, and if you watch Francesco de' Pazzi's video, it ends with a close-up of the slain Giovanni's face, the noose still around his neck.
Ezio slowly pushing his hidden blade into Checco Orsi's throat. It makes what Altair did to Majd Addin look friendly by comparison.
The Harlequin.
The Doctor for every courtesan in Roma, especially Fiora Cavazza who despite all the killings she participated in and madmen she encountered at this moment is scared out of her mind when she has to meet him for Cesare. Thankfully for her, she realizes one second before it's too late than he's a Dirty Coward.
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