Fig. 3

PDIA3P1 represses OCT4 degradation via the ubiquitin–proteasome pathway. A, B qRT-PCR analyses of OCT4 mRNA levels in cells with transfection with PDIA3P1-siRNA or PDIA3P1 expression plasmid (ns, no significance, student t-test, the data represent the mean ± S.D. of triplicates). C, D Western blot analysis of OCT4 protein stability using CHX (200 µg/mL) when PDIA3P1 was knocked down (C) or PDIA3P1 was overexpressed (D). E, F Western blot analysis of OCT4 in cells transfected with PDIA3P1 siRNA or expression plasmid in the presence of autophagy lysosomes inhibitor Chloroquine (CQ, 20 µM). G Treatment of the PDIA3P1-knockdown TE-1 and Eca-109 cells with the 26S protostome inhibitor MG132 (10 µM), western blot analysis expression of OCT4. H PDIA3P1 transduced KYSE-30 and KYSE-150 cells were treated with MG-132 (10 µM) and then the OCT4 expression level was examined by western blotting. I, J Western blot of the ubiquitination of OCT4 in cells that silenced PDIA3P1 or overexpressed PDIA3P1. IgG was used as a negative control