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https://www.aseprite.org/docs/color-mode/#indexed
"An important concept on Indexed images is that for transparent layers, we need a special index to act as the transparent color. Generally this index is 0, but you can change it from Sprite > Properties menu."
It means that transparent layers need one index that is used as transparency, only the background layer can use the transparent color as a solid color (Layer > Background From Layer to convert a transparent layer into a background)
I hope this help!
Maybe in the future, when we add mask layers this could be improved (e.g. transparent layers could use a mask layer for transparent pixels instead of a specific index as transparent)