Minor spoilers throughout.

Kris, the protagonist, has been they / themmed 49 times in the first 4 chapters, has never been referred to by any other pronoun and has a hidden sequence where they discover a childhood memory called “man country” where a sign says “and not-men are ok, too” and people (and by that, i mean cis people) still he / him them and insist that man country has nothing to do with transness. Oh, they also love dressing up in all kinds of glammy, genderfucky ways and look like this in the Dark World and the game starts with a big sequence where you get told that in this world, nobody gets to be themselves, your chosen name will be discarded and you are instead called Kris.

Ralsei, one of the other main characters, has presented like a femboy since the start of the game, wears pink eyeshadow and mascara and most recently dressed up as a princess in a pink gothic lolita dress. This, for reference, is Ralsei:

At the end of chapter 5, their pronouns magically changed to they / them after they have been told the entire chapter by an NPC who knows them from before the game that they “have some figuring out to do” right after they talked about a bunch of flowers who identify as humans, are kinda furry and talk about figuring out their gender a lot, which maaaaay be a trans allegory. The same NPC also gave him a pink and blue drink that apparently they had been staring at for several minutes, and previously you could equip pink, white and blue ribbons on them.

Oh and their central character arc is about them not allowing themselves to live their own life, to constantly exist for the benefit of others while denying themselves everything, and both this extreme people pleasing and the way they are constantly depressed and dissociated makes every transfem that knows the game immediately yell “one of us!”, as it is exactly the kind of unpersoned emptyness many of us have felt as eggs. Oh and there are tons of references to eggs around them, and they have a ton of stereotypical transfem kinks from being stepped on to being living furniture to petplay to suspension bondage. I think that’s most of the main stuff, this gay little goat is seriously the eggiest character i have ever seen in any piece of media, even moreso than pre-coming out Bridget from Guilty Gear. Like, it’s not just the way they present and act, they really capture the entire egg condition so well.

Noelle, the girl with the antlers in OOP, is mostly thought of as trans because her room is pink, white and blue and she is a female deer with antlers. But it has later been revealed that she is a reindeer, where both sexes grow antlers, so she might be cis. Regardless, she is extremely lesbian and has a heartwarming, wholesome yuri arc where she is the love interest of the last main character, Susie. Who is a gender noncomforming, black coded butch. You might begin to notice by now that this game is extremely queer. I haven’t even gotten into the other trans or trans coded characters, there are about 10 of them i think, but i guess you ca notice a certain pattern by now. The only people in this game who are possibly cishet are older NPCs that are usually the parents of other characters, and many of them are strongly hinted at to be bi.

afaik it’s all very subtle. Kris is NB and nobody questions it. Some of the ghosts seem to have flexible genders as well.