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Symbols
rising of the moon
There’s also the issue of the moon, as its appearance in the sky is the real signal that night has come and the men can stop waiting for Godot
tree
The tree is the only distinct piece of the setting,; Jesus was crucified on a cross, but that cross is sometimes referred to as a "tree," as in, "Jesus was nailed to the tree." That Vladimir and Estragon contemplate hanging themselves from the tree is likely a reference to the crucifixion