Fig 2: A landscape analysis of STAT5A gene mutations in ovarian cancer. (A) Lollipop charts of the mutated STAT5A gene. (B) Oncoplot displays the somatic landscape of OV TCGA cohorts. The genes and samples are sorted according to their mutation frequency and histology; Above the legend, the bar plot shows the number of mutations burdened.

Fig 3: Staphylococcus aureus infection suppresses the phosphorylation of NF-?B p65, STAT5, mTOR, and S6 at 8 h. (A) Phosphorylation of NF-?B p65 and STAT5 in S. aureus-invaded BMECs. (B) Nuclear localization of phosphorylated STAT5 in S. aureus-invaded BMECs. Representative confocal microscopy images of the level of p-STAT5 (Green) in cells that were co-stained with DAPI (blue). Scale bars represent 20 µm. (C) Nuclear localization of phosphorylated NF-?B p65 in S. aureus-invaded BMECs. Representative confocal microscopy images of the level of p-NF-?B p65 (Green) in cells that were co-stained with DAPI (blue). Scale bars represent 10 µm. (D) mTORC1 signaling in S. aureus-invaded BMECs. The resolved bands were quantified using Gel-Pro Analyzer 4.0 (Media Cybernetics, Inc., Rockville, MD, USA). *p < 0.05; **p < 0.01. n = 3 independent experiments.

Fig 4: C/EBPβ regulates the JAK/STAT signaling pathway in human TNBC. (A) Hierarchical cluster analysis of DEGs (|logFC| > 0.5 & P < 1E‐4) between C/EBPβ_high and C/EBPβ_low TNBC samples. Each cell in the matrix represents the expression of a gene in an individual sample. (B) Volcano plot of differential gene expression between C/EBPβ_high and C/EBPβ_low TNBC samples. Each dot represents one gene that had detectable expression in the RNA‐seq experiment. The horizontal dashed line marks the threshold (P.adj < 0.05) for defining a gene as significantly upregulated or downregulated in C/EBPβ_high samples compared with C/EBPβ_low samples. The vertical dashed lines represent twofold differences in expression. Significantly DEGs (|log2(FC)| > 1 and P.adj < 0.05) are marked in red. (C) Gene set enrichment analysis (GSEA) score curves. The JAK/STAT signaling pathway enrichment plot is shown. Black bars represent the position of genes of the JAK/STAT signaling pathway in the ranked list together with the running enrichment score (plotted in green). (D) The C/EBPβ binding motif was obtained from the JASPAR database. The character and size of each logo represent the proportion of a base at a specific site. (E) The positions of potential C/EBPβ‐binding sites at the STAT5A gene, defined by its binding motif, are shown. (F) C/EBPβ ChIP‐seq data (GSM2330565) were analyzed to determine C/EBPβ‐binding signals at the STAT5A gene. (G) qRT‐PCR assays were performed to examine the effect of C/EBPβ knockdown on JAK/STAT signaling pathway genes. Three independent experiments were carried out, and the results were represented as mean + S.D. *P < 0.05, **P < 0.01, ***P < 0.001. Unpaired t‐test. (H) Western blotting assay was carried out to confirm the change in STAT5A expression at the protein level upon C/EBPβ knockdown.

Fig 5: Evaluation of STAT5A expression in ovarian cancer tissues and cell lines. (A) Immunohistochemical staining of STAT5A and p-STAT5A on ovarian cancer and adjacent tissues from TMA samples (n=90). Black scale bar, 50 µm. (B) The ROC curves of STAT5A and P-STAT5A are based on independent tests. (C) Kaplan–Meier OS curves based on STAT5A or P-STAT5A level. (D) Immunoblots illustrating the basal expression of STAT5A, P-STAT5A (Tyr 694), MMP12, and MMP2 in the indicated human ovarian serous adenocarcinoma cell (HO8910) and normal ovarian epithelial cell line (IOSE80). (E, F) The mRNA and corresponding protein expression levels of SATAT5A were determined 48 h post-transfection with pcDNA-STAT5A plasmids in HO8910 cells. For normalization, GAPDH was used as an internal reference. MW: molecular weight. (G) Scratch-wound assay to quantify HO8910 cell migration ability. The scratches were recorded at 0 h and 48 h after scratching. Yellow lines indicated the scratched edges. White scale bar, 100 µm. (H) Transwell invasion assay with Matrigel. Cell invasion abilities were measured 48 h after cells or STAT5A overexpressing cells seeding onto a Matrigel-coated transwell filter. Black scale bar, 200 µm. **** P<0.0001, ns: not significantly determined by one-way ANOVA (E, F, H), Mann-Whitney test (B), Chi-square test, Kaplan-Meier survival analysis, and Log-rank statistical test (C).