Supplementary Figure 7: Extended data for Main Fig. 7.
From: An IRAK1–PIN1 signalling axis drives intrinsic tumour resistance to radiation therapy
(a) HeLa cells treated as in Fig. 7a analyzed by alamarBlue at 5 dpIR. Data expressed as means of 2 independent experiments in triplicates. CI, combination index, see (e). (b) Indicated cell lines treated and analyzed as in (a) with buparvaquone (0.5 µg/mL in BHY, 1 µg/mL in CAL27, SAS, and SW480) and R406 (0.1 µg/mL in BHY, 0.5 µg/mL in CAL27, SAS, and SW480). Data expressed as means of 2 independent experiments in triplicates. CI, see (g). (c-d) Clonogenic assays of CAL27 (c) and SAS (d) cells treated with subtherapeutic concentrations of the indicated drugs alone or in combination with IR (Gy); buparvaquone: 0.5 µg/mL (BHY), 1 µg/mL otherwise; R406: 0.1 µg/mL (BHY), 0.5 µg/mL otherwise. Data represented as means ± SD of n = 3 independent experiments in triplicates. (e-g) CI values for panel a (e), Fig. 7c (f) and panel b (g). (e) Each drug tested at two doses, thereby representing four data points after treatment with 0, 2.5, or 5 Gy IR. After 2.5 or 5 Gy IR, every combination of IRAK1+PIN1 inhibitor produced CI’s <0.8–1, indicating synergy. (f) CI values for p53MK/MK embryos treated with four possible combinations of two doses of EGCG and R406. At greater than 50% fractions affected, EGCG + R406 combination showed <0.8 CI, indicating synergy. Bottom-right data point is dose combination in Fig. 7c. (g) CI values for BHY, CAL27, SAS, and SW480 cells after indicated drug/IR combinations. Each data point represents mean of n = 3 independent experiments. (h) Extended data for Fig. 7e-g showing tumor volume (top) and animal weight (bottom) assays. Data expressed as means ± SEM, n = 5 mice per group. (i) Cells treated and analyzed as in (a), but substituting TP53 WT and null HCT116 cells for HeLa. Subtherapeutic concentrations as established for HCT116 cells were: R406, 0.1 μg/ml; Gin-Rb1, 5 μg/ml; buparvaquone 0.5 μg/ml; EGCG, 5 μg/ml. Data expressed as means of 2 independent experiments in triplicates. *P <0.05, **P <0.005, ***P <0.0005, two-tailed Student’s t-test. See Supplementary Table 4 for statistics source data including precise P values.
