The current (and unlikely to change) status of threadson win32 is that you basically can't perform gui operations from a subthread, even with gdk_threads_enter/leave. Essentially, the X11 and win32 gui/threading models are different, and making gtk work the same on both would be very hard.
Does gdk_pixbuf_new_from_file count as gui operation ?
Yes. To be safe, consider any gtk & gdk call to be a gui operation.
BTW, the underlying problem is fixable but it'll require a fair amount of work in the gdk implementation, see http://bugzilla.gnome.org/show_bug.cgi?id=60620
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