Figure 4.
USP9X regulates RIT1M90I abundance and proliferation of NCI-H2110 cells
(A) Western blot of NCI-H2110 cells treated with indicated siRNAs for 72 h. Vinculin serves as a loading control.
(B) Quantification of western blot bands based on (A) and additional replicates. Data shown are the mean ± SD of three independent experiments with 2–3 technical replicates per condition. p value calculated by paired two-tailed t test.
(C) Western blot of NCI-H2110iCas9 cells treated with 1 μg/mL Dox for 7 days to induce Cas9 expression. Cyclophilin A serves as a loading control.
(D) Quantification of Western blot bands in (C). Data shown are the mean ± s.d. of two independent experiments. p value calculated by paired two-tailed t test.
(E) Relative expression of RIT1 as determined by qPCR and ΔΔCt analysis. Data shown are the mean ± SD of three technical replicates per condition. Data are representative results from n = 2 independent experiments. ns = not significant by paired two-tailed t test.
(F) Western blot of NCI-H2110 cells transduced with two unique shRNAs targeting USP9X (shUSP9X #1 and shUSP9X #2). Vinculin serves as a loading control.
(G) Proliferation of parental NCI-H2110 cells and shUSP9X knockdown cell lines over time. p values calculated by unpaired two-tailed t-tests. ∗p = 0.01645; ∗∗p = 0.00245.