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Oh boy, here come the feminists.
It’s a reference to the episode “Magic Duel” where, spoiler alert!… Twilight pretends to change Applejack’s sex, then change her back! It then reveals to be a parlor trick, as it’s actually just Big Mac cross-dressing as AJ, and that the puff of smoke Twilight made was just to have the two switch places. Though the “eeyup” sort of gives the trick away, early…