Fig. 8. Degradation of E4B substrate proteins assayed by CHX chase.
(A) Comparison of the rate of substrate degradation in HEK293 cells with E4B overexpressed (E4B OE) or inhibition of E4B expression by shRNA (shE4B). A total of 5 × 106 cells were transfected with various plasmids, and the cells were treated with CHX (100 mg/ml) 48 hours after transfection. Cell extracts were collected at 0, 2, 4, and 6 hours after incubation with CHX, followed by immunoblotting with substrate-specific antibodies. (B) Quantitative analysis of the levels of the substrate proteins in the cells after CHX chase. Data are representative of three independent trials.