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. 2020 Feb 7;11(2):110. doi: 10.1038/s41419-020-2224-7

Fig. 4. DHODH inhibition induces cell cycle arrest in p53-deficient cells.

Fig. 4

a–g MDA-MB-231 cells were exposed to LFM (50 µM) for 72 h in the presence or absence of uridine (U; 50 µg/l). a Control and treated MDA-MB-231 cells as well as 4T1 parental, DHODH knock-out (KO) and DHODH reconstituted (rec.) cells were treated with nocodazole (10 µM, o/n) and analyzed for G2/M arrest using propidium iodide (2.5 µg/ml) staining detected by flow cytometry. b MDA-MB-231 and 4T1 cells were assessed for cell cycle distribution. The asterisk indicates significantly different values for S-phase changes. c Cyclin E and d Chk1 pS317, Chk1, p53 pS15, p53, p21, and H3 pS10 protein levels were detected by immunoblot. β-Actin and GAPDH were used as a loading control. e Accumulation of RPA32 protein in MDA-MB-231 cells was evaluated by immunofluorescence and f immunoblot. VDAC was used as a loading control. Scale bar represents 15 μm. g Accumulation of BrdU staining on non-denaturated ss-DNA in MDA-MB-231 cells was evaluated by FACS. In a, b, and g, data are shown as mean ± SEM, n = 3. *P < 0.05, two-way ANOVA. In other panels, representative experiment (from total number of three experiments) is shown.