Transparent background in a Tkinter window

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It is possible, but it’s OS-dependent. This will work in Windows:

import Tkinter as tk # Python 2
import tkinter as tk # Python 3
root = tk.Tk()
# The image must be stored to Tk or it will be garbage collected.
root.image = tk.PhotoImage(file="startup.gif")
label = tk.Label(root, image=root.image, bg='white')
root.overrideredirect(True)
root.geometry("+250+250")
root.lift()
root.wm_attributes("-topmost", True)
root.wm_attributes("-disabled", True)
root.wm_attributes("-transparentcolor", "white")
label.pack()
label.mainloop()

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