Sam Raimi's The Amazing Spider-Man franchise.
However, when both mainstream and comic fans think of Aunt May in live-action, it's likely Rosemary Harris' take on the character from Raimi's trilogy. Out of the three Mays that have been depicted on the big screen thus far, Harris is the one who, in spirit, resembled Peter's aunt from the comics. While she was a ing player in Tobey Maguire's series, her Aunt May had still so many iconic moments from all three films. May always had at least one motivational speech in every Spider-Man film, if not more as she would offer words of wisdom to her nephew.
While Peter never straight up told Aunt May that he was in fact Spider-Man, there is a very strong possibility that she already knows his secret. Even though Harris' Aunt May didn't have a big finding-out-scene as Tomai did in Uncle Ben died and he could have stopped it from happening. When Peter goes to see her, he witnesses Aunt May being in the middle of packing everything up as she had to move out.
As Aunt May forgives and thanks Peter for telling her the truth, it's what she says afterward that creates the possibility of her knowing that he's Spider-Man. A neighborhood kid Henry is helping her with the move and asks Peter where Spider-Man is, since this was not too long after Peter had given up being the web-slinger. As Peter sees Henry's disappointment, Aunt May also chimes in and wonders what has happened to the friendly neighborhood web-crawler. But it's how Aunt May describes what Spider-Man means to people where the hints start to come up. There are also specific parts in her speech that feel very directed towards Peter as opposed to Aunt May talking about it universally:
I believe there's a hero in all of us that keeps us honest, gives us strength, makes us noble, and finally allows us to die with pride. Even though sometimes we have to be steady and give up the thing we want the most. Even our dreams. Spider-Man did that for Henry and he wonders where he's gone. He needs him [emphasis added].
From how Harris delivers the speech and how she engages with Maguire, it's clear as day that she was telling Peter that the world needs him to be Spider-Man. Aunt May's reference to having to give up the thing they want the most, as well as their dreams, is an obvious nod to how Peter didn't allow himself to be with Mary Jane. It's understandable to then ask why Aunt May never said anything in Spider-Man 2 or Spider-Man 2.
