Fig. 1: JAK–STAT signaling is required for Enz resistance in TP53/RB1-deficient mCRPC.
a, Heat map representing the significantly changed signaling pathways in LNCaP/AR cell lines transduced with annotated shRNAs based on GSEA analysis. Three comparisons are presented. Reads from n = 3 independently treated cell cultures in each group were used for analysis. Signaling pathways concomitantly altered with TP53/RB1 loss and SOX2 upregulation are labeled with a red bracket. b, Relative gene expression of canonical genes activated in the JAK–STAT signaling pathway in LNCaP/AR cells transduced with Cas9 and annotated guide RNAs; n = 3 independently treated cell cultures. P values were calculated using a two-way ANOVA with a Bonferroni multiple-comparison test. c, Relative cell numbers of LNCaP/AR cells transduced with Cas9 and annotated CRISPR guide RNAs. Cells were treated with 10 µM Enz for 8 d, and cell numbers (viability) were measured using a CellTiter-Glo assay, with all values normalized to the sgTP53/RB1 group; n = 3 independently treated cell cultures. P values were calculated by one-way ANOVA with a Bonferroni multiple-comparison test; RLU, relative light units. d, Relative cell numbers of LNCaP/AR cells transduced with Cas9 and annotated CRISPR guide RNAs. Cells were treated with 10 µM Enz for 8 d, and cell numbers (viability) were measured using a CellTiter-Glo assay, with all values normalized to the sgTP53/RB1 group; n = 3 independently treated cell cultures. P values were calculated by one-way ANOVA with a Bonferroni multiple-comparison test; NS, not significant. e, Tumor growth curve of xenografted LNCaP/AR cells transduced with Cas9 and annotated guide RNAs in castrated mice. Cas denotes castration 2 weeks before grafting. Enz denotes Enz treatment at 10 mg kg–1 from day 1 of grafting; n = number of independent xenografted tumors in each group (two tumors per mouse); sgNT, n = 8 tumors; sgTP53/RB1, n = 12 tumors; sgTP53/RB1/JAK1, n = 8 tumors; sgTP53/RB1/STAT1, n = 12 tumors. P values were calculated by two-way ANOVA with a Bonferroni multiple-comparison test. f, IHC staining of JAK–STAT proteins on annotated xenografted tumor slides showing representative images of n = 2 independent tumors.
