For me, it was Princess Rosalina’s backstory in Super Mario Galaxy.
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The first time was in Mass Effect 3, when Liara shows you her project in your cabin. For me, this was the first time in the trilogy where I felt that we could really lose the war against the reapers.
Another very good moment was in Life is Strange Before the Storm, when Chloe finds her dad’s car in the junkyard. That hit hard.