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After 30 minutes, the media was replaced with fresh serum-free media and incubated for further 24 hours

Equal amounts of protein were loaded on SDSpolyacrylamide gel electrophoresis (SDSPAGE), using Mini-PROTEAN TGX gels (Bio-Rad, 4561096) and transferred to a 0.2-mm nitrocellulose membrane (Bio-Rad, 1620112)

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