Fig. 2
ZC3H18 depletion reduces BRCA1 mRNA and protein levels. a and b Indicated cell lines were transfected with control luciferase (Luc), ZC3H18 (ZC3), or BRCA1 siRNAs. Forty-eight hours after transfection, the cells were immunoblotted for the indicated antigens (a) and analyzed by qRT-PCR for ZC3H18 and BRCA1 mRNA levels, which are expressed relative to GAPDH mRNA levels as an internal control (b). c Rescue of BRCA1 mRNA and protein in ZC3H18-depleted OVCAR-8 cells with siRNA-resistant SFB-ZC3H18. OVCAR-8 cells were transfected with Luc or ZC3H18 siRNA plus empty vector (EV) or siRNA-resistant ZC3H18 plasmid (SFB-ZC3H18). BRCA1 mRNA and proteins levels were assessed by qRT-PCR (c) and immunoblotting (Supplementary Fig. 4b), respectively. d Ectopic BRCA1 expression rescues the HR defect caused by ZC3H18 depletion. OCVAR-8 DR-GFP cells were transfected with empty vector (EV) or HA-BRCA1-expressing plasmid with either control luciferase (Luc) or ZC3H18 (ZC3) siRNAs. Seventy-two hours later, the cells were analyzed for GFP by flow cytometry. Representative immunoblots in a are provided from three independent experiments. Unprocessed blots are provided in Source data file. Shown are means ± SEM from three independent experiments in b–d. *p < 0.05, **p < 0.01, ***p < 0.001, unpaired Student’s t-test
