Fig. 1: Emergence of CSCs following IFN-I treatment.
From: Type I IFNs promote cancer cell stemness by triggering the epigenetic regulator KDM1B
a, Multiparametric flow cytometry analysis of the illustrated CSC surface markers in MCA205 and AT3 cells treated with mock (CTR) or IFN-I (6 × 103 U ml–1, 72 h). Representative biparametric plots and histograms showing CD133+CD24+CD44+ percentages (mean ± s.e.m. with individual data point, n = 3 and n = 4 independent experiments) are shown. For more details on gating strategies, see Supplementary Fig. 1. b, Flow cytometry analyses of CD44L and CD44H percentages (top) and qRT–PCR analyses of the reported TF (bottom) in FACS-isolated CD133− and CD133+ MCA205 cells treated as in a. Mean ± s.e.m. with individual data point, n = 3 independent experiments. qRT–PCR data are reported as mean fold change (FC) ± s.e.m. over CTR after Ppia intrasample normalization, n = 3 and n = 2 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001; for exact P values, see Supplementary Table 1. c, SP (Hoechst 33342− within propidium iodide, PI−) in MCA205 and AT3 cells left untreated (black), treated with VRP (100 μM, light green), IFN-I (blue) or VRP + IFN-I (dark green). Mean ± s.e.m. with individual data point, n = 9 and n = 6 independent experiments. d, TF expression levels in IFN-I-treated MCA205 cells. Data are reported as in b, n = 3 and n = 4 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, see Supplementary Table 1 for exact P values. e, Clonogenicity of MCA205 and AT3 cells plated in soft-agar upon treatment as in a. The number (mean ± s.e.m. and individual data point) of biologically independent samples collected over three independent experiments is shown. f, Ex vivo flow cytometry of CD44L and CD44H cells within the CD45 negative (CD45−) fraction of MCA205 tumors from C57Bl/6J mice either treated with one single dose (1 × 105 U) or repeated doses (2 × 104 U) of IFN-I. Mean ± s.e.m. and individual data points for 10 mice per group from two experimental replicates. a,b,d Unpaired two-sided Student’s t-test and unpaired two-sided Student’s t-test with Welch’s correction compared with CTR. c,f, Brown–Forsythe test with Dunnet’s correction and ordinary one-way ANOVA test followed by Bonferroni’s correction. e, Unpaired two-sided Student’s t-test with Welch’s correction and two-tailed Mann–Whitney test compared with CTR.
