uS5 is required for the cellular accumulation of ZNF277 and PRMT3.
A, Western blotting analysis of the indicated proteins using extracts prepared from HEK 293 cells that stably express inducible version of a control shRNA (lanes 1 and 4) and uS5-specific shRNAs (lanes 2, 3, 5, and 6). The cells were grown in the absence (lanes 1–3) or presence (lanes 4–6) of doxycycline (Dox) to induce the expression of the different shRNAs. B and C, quantification of protein (B) and mRNA (C) levels for the indicated genes as determined by immunoblotting (B) and RT-qPCR (C), respectively, using extracts of control (shControl) and uS5-deficient (shuS5-1 and shuS5-10) cells cultured in the presence of doxycycline. Protein levels (B) were normalized to tubulin and expressed relative to shControl cells, whereas mRNA levels (C) were normalized to the GAPDH mRNA and expressed relative shControl cells. The data and error bars represent the averages and standard deviation from at least three independent experiments. **, p ≤ 0.01; ****, p ≤ 0.0001; Student's t test.