Fig. 7.

TMEM129 and Ube2J2 are required for US11-induced FcRn ubiquitination, a, b TMEM129 and Ube2J2 is required for US11-induced FcRn ubiquitination. HeLa^FcRn+US11 cells were transfected with 20 nM TMEM129, Ube2J1, or Ube2J2 siRNA oligomers for 48 h or empty vector. Cells were subsequently treated with 50 μM MG132 for 4 h and then lysed. All immunoprecipitates or cell lysates (20 μg) as internal controls were subject to western blotting with corresponding Abs as indicated. c, d HeLa^US11+FcRn cells were transfected with 20 nM Ube2J1 (c, top) and Ube2J2 (c, bottom) siRNA oligomers for 48 h. Cells were then treated with CHX (100 μg/ml) and chased for the indicated time. The cells were lysed after CHX treatment and cell lysates (20 μg) were subject to western blotting with Abs as indicated. The level of remaining FcRn (d) in Ube2j1 siRNA-treated cells (black) or Ube2j2 siRNA-treated cells (red) was quantified as the percentage of the β-tubulin level at different time points. The percentage of time point 0 (min) is assigned a value of 100% and the values from other time points are normalized to this value. Error bars represented mean ± SEM of three independent repeats