EBNA3C Augments Pim-1 Mediated Phosphorylation and Degradation of p21 to Promote B-Cell Proliferation
Figure 5
EBNA3C contributes to Pim-1 stabilization.
A) 10 million HEK-293 cells were co-transfected with Myc-Pim-1 and either vector control (lanes 1 and 3) or Flag-EBNA3C (lanes 2 and 4) expression constructs. After 36 hrs of post-transfection, transfected cells were treated with either 20 µM MG132 (+ lanes) or DMSO (− lanes) for additional 6 hrs and cell lysates were resolved by 10% SDS-PAGE and Western blot was performed with the indicated antibodies. B) HEK-293 cells were transfected with above mentioned expression vectors and At 36 hrs of post-transfection, cells were treated with 40 µg/ml cyclohexamide (CHX) for 0, 3, 6 hrs. Cells were lysed and protein samples were resolved by 10% SDS-PAGE. Western blot was performed by specific antibodies shown. C–D) BJAB, BJAB10, LCL1, sh-Ctrl and sh-EBNA3C cells were treated with 40 µg/ml cyclohexamide (CHX) for indicated time periods. Cell lysates were resolved by 10% SDS-PAGE. Western blot analysis was performed with indicated antibodies. GAPDH blot was shown for internal loading control.