Supplementary Figure 1: Extended data for Main Fig. 1.
From: An IRAK1–PIN1 signalling axis drives intrinsic tumour resistance to radiation therapy
(a) Representative images of 25 hpf embryos stained with AO, with conditions indicated, 7 hpIR (15 Gy Cs γ-IR TBI delivered at 18 hpf). Scale bar, 0.5 mm. (b) Quantification of AO positivity in the neural tubes of embryos from panel (a). Number of quantified embryos were: p53+/+, DMSO-treated: n of non-irradiated = 7, n of irradiated = 7; p53+/+, Gö6976-treated: n of non-irradiated = 6, n of irradiated = 8. p53MK/MK, DMSO-treated: n of non-irradiated = 7, n of irradiated = 8; p53MK/MK, Gö6976-treated: n of non-irradiated = 7, n of irradiated = 8; altogether obtained from 2 independent experiments. Data expressed as means ± SD, ***P < 0.0001, n.s., not significant, two-tailed Student’s t-test. See Supplementary Table 4 for statistics source data including precise P values. (c) Schematic of primary screen. Embryos arrayed in multi-well plates and treated with drug libraries at 17 hpf, irradiated (15 Gy) at 18 hpf, drug-treated for 6 hr, washout at 24 hpf, and scored for DTCs at 120 hpf. (d) Primary screen results. 640 FDA-approved drugs (X-axis, 2 μg/mL each) scored for DTC penetrance (Y-axis) in >12 embryos/drug. Gö6976, used as positive control, labeled in red. Drugs producing >75% DTC were considered primary hits and carried over to secondary screen. (e) Secondary screen. For each primary hit in (d), embryos were arrayed in a similar schema but half of embryos were non-irradiated. Results plotted in (f-g) below. (f) Schematic of 136 drugs screened in secondary screen (e). Quadrants determined based on greater than or less than 50% potency (%DTC +IR) and greater or less than a selectivity ratio [(%DTC +IR)/(%DTC –IR)] of 4:1. Quadrant I: selective but not potent; II: both potent and selective; III: potent but not selective; IV: neither potent nor selective. Identity of each compound in the screen described in Supplementary Tables 1-2. (g) Each drug was investigated by SEA for possibly targeting IRAK1/4 or PIN1. Red, predicted to bind IRAK1/4; blue, predicted to bind PIN1/4; purple, predicted to bind both IRAK1/4 and PIN1/4. Additional, possibly lethal off-targets were not investigated.
