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The death of Bambi's mother. It traumatized a lot of little kids that saw that movie. Steven Spielberg even recalled in his childhood waking up in the middle of the night and going to his parents' room to check if they were both still alive.
The death of Bambi's mom is one of the most famous tear jerkers in the history of film, to the point it's been parodied relentlessly in the decades since its release (q.v. the Animaniacs episode "Bumbie's Mom"). Watching the scene in context, seeing his mother give up her own life just for Bambi to survive, as well as Bambi being told by the Great Prince what happened to her is the cinematic equivalent of a punch to the gut.