Fig. 4

USP9X regulates EGLN3 at level of protein post translational modification. A HEK293T cells transiently co-expressing HA-EGLN3 and pCDH, Flag-USP9X WT or FLAG-USP9X C1566S were subjected to analyze by immunoblotting using indicated antibody. B RBE cells transiently expressing pCDH, Flag-USP9X WT or FLAG-USP9X C1566S were subjected to analyze by immunoblotting using indicated antibody. C RBE cells transiently expressing pCDH or Flag-USP9X WT were added with DMSO or WP1130 then subjected to analyze by immunoblotting using indicated antibody. D RBE (left) and HUCCT (right) cells stably expressing shNC, shUSP9X #1 and shUSP9X #2 were analyzed by real-time PCR. E RBE (left) and HUCCT (right) cells stably expressing pCDH and Flag-RNF216 were analyzed by real-time PCR. F RBE (left) and HUCCT (right) cells added with DMSO or WP1130 were analyzed by real-time PCR. G RBE (left) and HUCCT (right) cells stably expressing shNC and shRNF216 #1 were subjected to immunoblotting analysis with the indicated antibodies after 48 h of transfection. Cells were treated with 20 μM MG-132 for 6 h before harvest