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In [[5.09 The Real Ghostbusters]] when Dean and Sam are trying to convince a girl to act as bait for the ghosts, Dean says to Sam "Just give her the puppy dog thing, okay?"
In [[10.02 Reichenbach]] Sam employs the puppy dog eyes, but due to his corruption from the [[Mark of Cain]], Dean ridicules Sam for using them.
In [[15.10 The Heroes' Journey]], its shown that the puppy dog eyes no longer work after [[God]] has downgraded the Winchesters to normal people again. The [[werewolf]] Brad comments on Sam's puppy dog eyes and asks if it ever actually works when Sam tries it on him.
