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Darkiplier, Warfstache, and Yancy (along with another groovy musical number) will all make appearances in Part 2.
The voice behind the door teased the appearances of Dark and Yancy, and an easter egg shows Wilford disco dancing, revealing that his proper appearance is in part 2.

Old Mark sabotaged the ship.
Old Mark can be seen running around in the background of the first video. It’s noted that some of the ship’s damage looks like sabotage—for example, the turrets beginning to attack the crew when Mark states that should be impossible since he programmed them. He could also easily change that programming, and as the Head Engineer he’s one of the two people who could change other things on the ship. Old Mark is sabotaging things in a misguided attempt to stop the Captain and his past self, thereby causing the paradox that he’s trying to stop.
  • Confirmed.

The Invincible I was shown in A Heist with Markiplier
A spaceship is shown on the background in Ending 30 as you float in space with Mark. Many suggested that this belongs to the two-headed alien, but when you meet him, you don't appear in a spaceship.

Meta WMG: In Space, Heist, and the projects past and future, are all connected, but not quite in the way we expect.
In Heist, one ending leads directly into A Date with Markiplier, with no other endings having any apparent Sequel Hook to a past Markiplier adventure. Here, in In Space, the crystal from Heist is implied to have come from the warp core Mark built, with a link card taking the viewer to Heist in the Golden Ending.With the possible exception of Who Killed Markiplier?, which may actually be chronologically the earliest story in the lot, each new installment takes place before the last.That, or they have a looser grip on causality than Doctor Who. Which is perfectly possible, especially given all the wormhole shenanigans this time around.

Simulation.
Seriously, why was this not the first WMG on this page?The digital glitches. The many incarnations of the same characters, sometimes replacing each other during particularly critical glitches. The universe literally rebooting from a 1980's OS.

Everyone is stuck in a series of computer simulations, their wetware time-shared between different programs to create simulacrums that play out each scenario. Who Killed Markiplier, A Date With Markiplier, A Heist With Markiplier, Wilford Motherloving Warfstache, Damien, and In Space With Markiplier with all its sub-stories are all playing at the same time for a worldwide audience of players. Except this last update had a major bug: When Markiplier sent the Warp Crystal back in time, he was forcing the thing to play the entirety of every simulation, which would include the simulation of the thing to play the entirety of every simulation, ad infinitum, causing the Captain and Markiplier to recurse through each virtual machine's simulation until the order is overridden (by not sending the thing 'back in time') or the system completely shut down from the strain. But by overriding the command, the innermost nested simulation played the contents of the thing instead of safely deleting it, allowing Darkiplier to get a DRM-free copy of the MacGuffin, just as planned.

Dorene is Darkiplier
It’s strongly implied that Dorene is Celine, but she can’t be just Celine. Her body was destroyed during the events of Who Killed Markiplier and it’s emphasized in Damien that she is just as much a part of Dark as Damien is, her mind was just asleep/in stasis. Their souls shouldn’t be able to exist individually anymore. So maybe this is what it looks like when she wakes up.As for why she’s here? Possibly to watch over the Captain with more subtlety than usual. Dorene is clearly an alias for her, and it sounds more like Dark’s name then her original name. She also demonstrates weird glitchy powers that look much like what Dark can do. Dark does show up in part 2 (not at the same time as Dorene), but you can see in Date that he’s capable of turning off the chromatic aberration, so who’s to say his weird powers don’t include shapeshifting?

The canon year that the series takes place in is 2081.
Assuming that the characters are the same age as their actors, adding their ages to the characters birth years (as seen here) causes them all to add up to 2081. This is likely the actual year that ISWM is supposed to take place in.

The wormhole bridges every universe, including ones with more meta-awareness than the one we started in.
In the "10 Years of Markiplier" livestream, Markiplier clarified several things.note 1. Mark the Head Engineer is not Actor Mark, but maybe Actor Mark could be Engineer Mark;2. In Space with Markiplier opens doors backward through time, forming nebulous links to previous stories and3. The wormhole drags every universe into itself like a funnel, where the multiverse collapses in on itself, and at the bottom of it is Mark, the Engineer, and you, the Captain.

The big take-away this troper had was this: while everyone exists in almost every universe, we are fundamentally different people in some timelines. For example, a version of Mark exists in every universe, but time and time again the Captain encounters Marks who have nothing in common with Engineer Mark beyond name and likeness. Sometimes he's an investigative engineer. Sometimes he's a camp counselor, or a renowned criminal, or a YouTuber. Maybe even an actor on parole.

This is why the Captain encounters Actor Mark under certain conditions. It also explains why there were references to In Space with Markiplier in Mark's regular videos between parts 1 and 2 (ignoring the more likely reason that he was simply plugging the show.) The universe our real world resides in wasn't quite in the wormhole's sphere of influence, but just close enough for it to bleed through in We Went Back, Drive Time Radio and Mark Is Too Stupid to Understand Sports. If you wanted to get really meta, you could argue the only reason it grazed our universe to begin with was its connection to the real-world Mark; not the same Mark at the center of the wormhole, but similar enough. An equivalent to him. There is no such thing as fiction, only non-fiction in the wrong universe.

The Invincible I never existed; Mark named the current ship Invincible II because he vaguely remembers the events of the Time Loop on a subconscious level.
Throughout the series, Mark repeatedly experiences Déjà Vu as events occur, with the implication that they've already occurred in alternate timelines of which he now has a vague awareness thanks to wormhole shenanigans. His memory of the first Invincible is similarly unreliable; at times he claims to recall her fate with perfect clarity, only to realize (often mid-sentence) that he doesn't actually remember — for example, this comment to Lady:
"You should've seen what happened to the first Invincible! ...I mean, actually, now that I think about it, I don't think anyone saw what happened to the first Invincible..." [looks at the Captain with a confused expression]

With the big reveal in Part 2 — that the story is a Time Loop, with the Captain letting Mark go back in time resulting in the whole thing starting over — we can infer that there never was an Invincible I, and that Mark subconsciously remembers something bad happened to a ship called Invincible, resulting in him naming the new ship Invincible II. In the Golden Ending, the ship — the same ship you commanded in Part 1, with the same crew — is now simply called Invincible, indicating that the time loop has broken and Mark was not influenced by déjà vu when he named it.

This is also supported by Mark's stealth ad for the series at the end of his playthrough of "Drive Time Radio", in which he claims that the Invincible II is humanity's first starship. Not the first successful starship, the first, period.

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