Fig. 3: RPS3 ubiquitination at K214 promotes dissociation of stalled 40S subunits. | Nature Communications

Fig. 3: RPS3 ubiquitination at K214 promotes dissociation of stalled 40S subunits.

From: E3 ubiquitin ligase RNF10 promotes dissociation of stalled ribosomes and responds to ribosomal subunit imbalance

Fig. 3

a FLAG-RPS3-WT and FLAG-RPS3-K214A were stably expressed in HeLa cells, and Ub1-RPS3, ± ANI treatment (2 h, 0.1 µg/ml), was assessed by Western blot analysis; Tubulin serves as loading control. The blot is representative of 3 independent experiments. b Polysome profile analysis of cells as in panel (a) after KD of RPS3 (siRNA S292) on day 1 followed by a second KD of RPS3 on day 3. Polysome profiles were recorded on day 5 after intermediate-dose ANI treatment (2 h, 0.1 µg/ml) or on day 6 after low-dose ANI treatment (for 48 h from day 4–6, 0.02 µg/ml). c Free 40S subunits and ribosomal half-mers (2.5 mer / disome) were quantified from polysome profiles as in panel (b) by calculating the ratio of the corresponding areas under the curve; shown are mean values ± SD (n = 4 biological replicates; p-values determined by two-tailed, paired t-test). Source data are provided as a Source Data file.

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