Mega 2560 Rev. 3: Not Working Without Initial USB Connection

Your posts are confusing.

You talk about external 9volt and USB, and then you talk about 5volt and USB.
And you didn't mention where you have connected external supply to.

A Mega needs an absolute minimum of 6.6volt on the V-in pin, or 7.3volt on the DC socket (including dips).
5volt external can be connected to the 5volt pin, but that could give unexpected results in combination with USB supply.

A sketch is saved to flash during upload, and does NOT change until you upload a new sketch.
A bad supply could reset/corrupt the Mega during bootup, but that does not affect the stored sketch.

Not sure what else you have attached to the Mega's 5volt pin.
I hope not the 152 addressable LEDs you mentioned in another thread.
Post a connection diagram, and exact specs of the supply.
Leo..