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. 2021 Oct 5;29(3):687–696. doi: 10.1038/s41418-021-00882-0

Fig. 3. WDR75 knockdown activates p53 and p21 and impairs proliferation.

Fig. 3

A U2OS cells were transfected with indicated siRNAs, cultivated for 3 days and cell lysates analyzed by immunoblotting with indicated antibodies. B U2OS cells were transfected with indicated siRNAs, next day 100,000 cells plated into six-well plates, incubated for indicated additional number of days and counted. Data shown as mean ± SD of three independent experiments (n = 3), each based on triplicate wells. Significance was determined by two-tailed t test: *P < 0.05. C U2OS cells were transfected with indicated siRNAs, cultivated for 3 days and cell cycle profiles analyzed by FACS. D U2OS cells were transfected with indicated siRNAs, cultivated for 3 days, treated with 10 EdU for 30 min and stained for incorporated EdU by click chemistry. Bar, 10 μM. E Quantification of EdU positive cells from D. Data shown as mean ± SD of three independent experiments (n = 3). Significance was determined by two-tailed t test: *P < 0.05. F U2OS cells were transfected with indicated siRNAs, incubated for 3 days, labeled with CldU/IdU and fork rate determined by DNA combing. Fork rates based on IdU track length are shown as box plot. Scored forks: 208 and 113 for siCON and siWDR75, respectively. Data shown is representative of two independent experiments.