Place it right after the scare is revealed, then follow it with the Vine Boom sound for a bigger hit, or lead into it with the Dun Dun Dun sound to build tension first.
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The clip is a short fearful emote, lasting just over a second, with a villager letting out a quick, high-pitched startled squeak. It feels nervously cute rather than truly scary, like a tiny cartoon panic that makes a jump scare feel playful. It comes from Animal Crossing, where villagers use the emote to react to something frightening.