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. Author manuscript; available in PMC: 2022 May 22.
Published in final edited form as: Nat Genet. 2021 May 10;53(6):881–894. doi: 10.1038/s41588-021-00859-2

Fig. 5 ∣. Sox2 overexpression confers enhanced dependency on Adar1 and promotes IFN pathway activation.

Fig. 5 ∣

a, Plot shows differential gene dependencies between SOX2-amplified and nonamplified cancer cell lines from CRISPR screening data. The effect size denotes mean difference of gene dependency score between the two groups with P values estimated from empirical Bayes moderated t-statistics using the Limma R package. b, Cell viability of human ESCC cell lines TT and KYSE70 is shown 5 d after shRNA-mediated ADAR1 silencing. Data were normalized to nontargeting shRNA control (n = 3 independent replicates) and are shown as mean ± s.d.; P values were calculated by two-sided one-way ANOVA with Benjamini–Hochberg correction. c, Growth curve for KYSE70 xenograft tumors (n = 6–8) with shRNA-mediated ADAR1 silencing. Data are shown as mean ± s.e.m.; ****P < 0.0001 calculated by two-way ANOVA with Benjamini–Hochberg correction. d, Cell viability of CPP (left) and SCPP (right) organoids is depicted following control or Adar1 silencing by shRNA (n = 3 independent replicates). Data are shown as mean ± s.d.; ****P < 0.0001 from two-way ANOVA with Benjamini–Hochberg correction. Technical replicates from one representative experiment are shown (n = 3). e, mRNA expression of ISGs (Mx1, Cxcl10 and Isg15) in CPP and SCPP organoids was assessed by RT–qPCR after control or Adar1 knockout (n = 3 independent replicates). Data are shown as mean ± s.d.; P values were calculated by two-sided unpaired t-tests. f, Cell viability of CPP and SCPP was assessed after transfection with dsRNA poly(I:C) or dsDNA poly(dA:dT). Data were normalized to mock infection control (n = 3 independent replicates). Data are mean ± s.d.; NS, not significant calculated by two-sided unpaired t-tests. g, mRNA expression of ISGs (Mx1, Cxcl10 and Isg15) in CPP and SCPP organoids was assessed by RT–qPCR 24 h after transfection with dsRNA poly(I:C) or dsDNA poly(dA:dT) (n = 3 independent replicates). Data are mean ± s.d.; NS, not significant; P values calculated by two-sided unpaired t-tests. h, Representative double immunofluorescence images in CPP and SCPP organoids with fluorescein isothiocyanate (FITC) (green), phalloidin (red) and 4′,6-diamidino-2-phenylindole (DAPI) (blue) were used to detect J2, Adar1 and nucleus, respectively. Scale bar, 80 μm or 30 μm. (This experiment was repeated once with similar results.) DMSO, dimethylsulfoxide; Dox, doxycycline.