The FBS used to culture cells destined for exosome experiments was depleted of exosomes by centrifugation at 140,000g for 3 hours; the supernatant of the centrifuged FBS was then used for cell culture. HT-29 cells were treated with DMSO or TSZ for 6 hours. The culture medium of the cells was collected and centrifuged at 1000g for 10 min, the supernatant of which was then centrifuged again at 10,000g for 30 min. The resulting supernatant was centrifuged again at 140,000g for 3 hours. The pellets (P140) were directly lysed in 1× SDS loading buffer and analyzed by Western blotting.

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