Fig. 6

RNF157 mediates K48 ubiquitination of DDX58. A HCCLM3 and Hep-3B cells were knocked down, Huh7 cells were overexpressed, and Western blot analysis was performed using RNF157 and DDX58 antibodies. Huh7-overexpressing cells were subjected to Western blot analysis with an RNF157 antibody and a DDX58 antibody. B HEK293T cells were cotransfected with different doses (0, 1, 2, or 4 μg) of HA-RNF157 and 4 μg of the Myc-DDX58 plasmid and cultured for 48 h. Western blot analysis was performed after treatment with MG132 (20 μM) for 6 h. C Lysates from HEK293T cells transiently cotransfected with Flag-Ub (K48, K63, KWT), HA-RNF157, or Myc-DDX58 were treated, immunoprecipitated with anti-Myc antibodies, and then treated with anti-HA, anti-Myc, or anti-Flag antibodies for Western blot analysis