Fig. 10. RNF113A promotes resistance to chemotherapy.

a Generation of Cisplatin-resistant lung cancer A549 cells (CR (4.5)). Pictures of both parental and resistant cells and WB analyses are shown. On the right, sensitivity of parental and resistant cells to 72 hours of treatment with Cisplatin. The percentage of viable cells in untreated parental or resistant cells was set to 100%. Data from two independent experiments in triplicates are shown (mean ± SD, ***p < 0.001, **p < 0.01, *p < 0.05, Student t-test). b RNF113A deficiency in CR (4.5) cells enhances cell death upon Cisplatin stimulation. On the left, cell survival upon Cisplatin treatment in resistant control and RNF113A-depleted A549 cells was assessed by FACS. The percentage of cells in early or late apoptosis is mentioned. On the right, FACS data from two independent experiments are illustrated in the histogram (Student t-test, ***p < 0.001). c RNF113A deficiency in Cisplatin-resistant lung cancer cells enhances DNA-PKcs phosphorylation upon DNA damage. Control or RNF113A-depleted Cisplatin-resistant A549 cells were treated or not with Cisplatin and WB analyses are shown. d RNF113A promotes SF3B2, Rad51 and RNF8 expression. Control or RNF113A-depleted Cisplatin-resistant A549 cells were untreated or stimulated with Cisplatin and WB analyses are shown. e RNF113A deficiency sensitizes Cisplatin-resistant lung cancer cells to BCL-2 inhibition. Control or RNF113A-depleted Cisplatin-resistant A549 cells were untreated or incubated with ABT737. FACS analyses were done to quantify cells undergoing early or late apoptosis. On the right, FACS data from two independent experiments are illustrated in the histogram (Student t-test, ***p < 0.001). f RNF113A deficiency sensitizes resistant lung cancer cells to Cisplatin-dependent cell death. Control or RNF113A-depleted Cisplatin-resistant A549 cells were transplanted into immunodeficient mice. Tumors were grown up to 0.1–0.2 mm³ and mice were treated with Cisplatin (1 mg/kg) six times every 3 days. Seven mice were used per experimental conditions. WB analyses were done with tumors generated from control (« C ») and RNF113A-depleted (« D ») Cisplatin-resistant cells. At the bottom, illustration of the size of tumors obtained after Cisplatin administration and quantification of tumor weights after Cisplatin administration (two experiments, Student t-test, **p < 0.01).