Fig. 5: Foxf2 suppresses Cxcl5 directly and indirectly.

Heatmap of RNA-Seq analysis of control and Foxf2-expressing WPMY-1 (N = 3 each) (a), in vitro cultured control and Foxf2-expressing mouse prostate stromal cells (N = 3 each) (b), and FACS-isolated control (N = 3) and Foxf2-expressing (N = 4) mouse prostate stromal cells from in vivo RM-1 tumors (b). Heatmaps depict fold changes in experiment versus control. Each gene is centered on average of control. c, d Gene ontology analyses of RNA-Seq analyses. e CAF gene signature scoring comparing Foxf2 overexpression (OE) with control WPMY-1 cells. Bars represent standard error. P-values by two-sided t-test. f Western blot analysis using control and Foxf2-expressing mouse prostate stromal cells. LPS: lipopolysaccharide. Experiments were repeated for three times with similar observation. qRT-PCR of Cxcl5 in control and Foxf2-expressing mouse prostate stromal cells transduced with and without super IκBα (g) or dominant negative STAT3 (dnSTAT3) (h). Dot plots show means ± s.d. of Cxcl5 expression from 3 independent experiments. Two-way ANOVA with Tukey’s multiple comparison test (i) ChIP analysis of Foxf2 binding at promoter of Cxcl5 in mouse prostate stromal cells. TSS: transcription start site. Loci at Ch.15 and Pdgfrα serve as negative and positive binding controls, respectively. Dot plot shows means ± s.d. of relative enrichment from 3 independent experiments. Two-sided unpaired t-test. Luciferase reporter assays determine activity of Cxcl5 reporter with and without mutations in putative Foxf2 (j) or NF-κB (k) binding sites in control and Foxf2-expressing mouse prostate stromal cells. Data represent means ± s.d. from 3 (j)−6(k) experiments. Two-sided unpaired t-test. Source data are provided as a Source Data file.