Fig. 7.

P-S134-GR promotes the expression of a 24-gene signature that correlated with poor prognosis in BC. a Venn diagram showing genes that are upregulated by TGFβ1 in MDA-MB-231 cells expressing either wt-GR or S134A-GR. The cutoffs used to define upregulation were a Benjamini-Hochberg value of less than 0.05 and a log2 fold-change of at least 1.5. b Supervised clustering of the 39 genes significantly upregulated in wt-GR cells by TGFβ1. cLEFTY2 and PIK3IP1 mRNA levels were assessed by qRT-PCR following normalization to Actin. Wt-GR and S134A-GR MDA-MB-231 cells were treated with either vehicle or TGFβ1 (10 ng/mL) for 6 h. Mean expression of three independent replicates ± SD is shown. Statistical significance was assessed by two-way ANOVA and Tukey post hoc for comparison within groups (*, p < 0.05). d Either wt-GR or S134A-GR MDA-MB-231 cells were treated with either vehicle or TGFβ1 (10 ng/mL) for 1 h. ChIP assays for the LEFTY2 and PIK3IP1 promoters to evaluate recruitment of GR were performed. The mean of three independent biological replicates are shown ± SD. Statistical significance was assessed by two-way ANOVA and Tukey post hoc for comparison within groups (*, p < 0.05; ****, p < 0.0001). e Kaplan-Meier curves are shown for the METABRIC dataset (n = 1904). Patients were separated by calculating the median expression of the previously identified GR Ser134-dependent 24-gene signature. This analysis was limited to 5 years of survival data. The results are significant with a logrank p value of p = .0008. f SurvExpress [51] was used to stratify TCGA breast cancer cohort based on their median prognostic index as determined by SurvExpress with the TGFβ1 pS134-GR 24-gene signature. The results were analyzed for significance with a logrank p value of p = .0030