Fig. 1: RPS3 monoubiquitination by RNF10 upon translation elongation stalling.
a Following control KD, KD of ZNF598 (S193) or KD of RNF10 (S214) for 72 h, HeLa cells were treated for 2 h with translation elongation inhibitors anisomycin (ANI, 0.1 µg/ml), cycloheximide (CHX, 100 µg/ml), or blasticidine (BLA, 100 µg/ml). RPS3 monoubiquitination (Ub1) was assessed by Western blot analysis. b Western blot analysis showing the effect of ANI treatment (2 h, 0.1 µg/ml) on Ub1-RPS3 in RNF10-KO HeLa cells (clones 81-7 and 29-7) compared to parental HeLa cells or a KO-control clone. The blot is representative of 3 independent experiments. c Polysome profile and fractionation analysis of HeLa RNF10-KO cell pools; the distribution of RNF10 and Ub1-RPS3 in the absence and presence of ANI treatment (2 h, 0.1 µg/ml) was assessed by Western blot analysis. Sharp peaks pointing downwards are artefacts of the electric signal of the fractionator. d The Ub1-RPS3 / RPS3 ratio (n = 1 biological replicate) in parental and KO-control HeLa cells in the presence of ANI was calculated by quantification of Western blots in panel (c). Source data are provided as a Source Data file.
