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Rosalina makes two apperances in FNaW: Origins, once as a easter egg, and another in a cutscene.

The Easter Egg can only occur on Night 3, where if you click a button on the computer several times a silhouette of Rosalina will appear.
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Rosalina in the Easter Egg.

[SPOILERS] Rosalina also appears in the Phone Call cutscene before Night 5, where she gets encountered by Waluigi, Peach, Mario, Luigi, and Wario (in his plumber outfit). Wario attacks her, and in response Rosalina opens up a portal that sucks up the souls from all 5 of their bodies, and the image of a stopwatch rewinding is shown.

Trivia

  • Rosalina is the only character from the Mario Universe not to appear as an enemy in any way, shape, or form in the FNaW universe.
    • [SPOILERS] Similarly, she's the only character from the pixel-style cutscenes in FNaW: Origins that Wario encounters but doesn't turn into a possessed spirit.
  • [SPOILERS] as revealed in the Full Story, Rosalina is sending the spirits of Wario and his gang back in time, since she knows that they'll keep killing people if they're not stopped.
    • [SPOILERS] On a similar note, the time warp featured in the pixel cutscene supposedly takes place after the Bad Ending of FNaW3, due to Wario wearing his plumber outfit and having a grey-colored head, similar to his skull-like head from FNaW3.