Fig 2: FGF10 and JAK1 can serve as independent predictive biomarkers. (a) Representative IHC images of EOC patient tissue stained for JAK1 or FGF10. IRS was used to attribute specimen according to their protein expression level for the Kaplan–Meier survival analysis shown in (b). Tissue samples with low (IRS = 0) or high (IRS ≥ 1) protein expression levels are shown (scale bar = 100 μm). (b) Kaplan–Meier survival analysis of progression-free survival (PFS) of 365 patients with high-grade serous EOC with low or high JAK1 or FGF10 expression level. Median survival of each group is indicated. Statistical significance was determined by Chi-Square statistics of the Log-Rank test (Mantel–Cox).

Fig 3: JAK1 and FGF10 acquire H3K27ac during cPt resistance development and are early and late markers for EOC development. (a,b) ChIP-seq tracks of H3K27ac intensity at the JAK1 (A) and FGF10 (B) gene in sensitive A2780 and resistant A2780cis cells. (c,d) Gene expression of JAK1 (c) and FGF10 (b) following cPt treatment (RT-qPCR) relative to SDHA control, normalized to untreated A2780 cells (0 h cPt). (e,f) H3K27ac signal at JAK1 (e) and FGF10 (f) following cPt treatment (ChIP-qPCR, relative to spike-in) normalized to untreated A2780 cells (0h cPt). (c–f) A2780 cells were treated with cPt (1 µM) for the indicated time points; 26d res and 42d res indicate cells which acquired resistance by treatment of A2780 cells with cPt (1 µM) for the indicated days. Mean ± SD, n = 3. (g) Analysis of JAK1 and FGF10 expression (RNA-seq) in patients with different FIGO stages of EOC from the GDC TCGA Ovarian Cancer (OV) cohort. Box plot with 2.5 and 97.5 percentile is shown for the indicated number of patient samples per group.

Fig 4: Induction of interferon (IFN) signaling by IFIT3. (A, B) HDF, HDF-Empty and HDF-IFIT3 cell extracts were analyzed by Western blotting using antibodies against (A) IFIT1, IFIT2 and IFIT3, (B) total STAT1 and phosho-STAT1 (Y701). The bands near the bottom of the gel in (A), IFIT2 and IFIT3, are nonspecific, cross-reactive proteins. (C) RNA was isolated from HDF, HDF-Empty and HDF-IFIT3 cells and IFNβ and specific interferon-stimulated genes (ISGs) (IFIT1, ISG15, OAS3, and MX1) mRNA levels were quantified by RT-qPCR. The results were normalized to glyceraldehyde-3- phosphate dehydrogenase (GAPDH) mRNA levels and fold change in HDF-IFIT3 cells was compared to HDF and HDF-Empty cell lines, and plotted as mean ± SD, n = 3. (D) HDF, HDF-Empty and HDF-IFIT3 cell extracts were analyzed by Western blotting for total TBK1, phospho-TBK1 (S172), total IRF3, and phopsho-IRF3 (S396). Human THP1 monocytes were left untreated or treated with IFN-stimulatory DNA (ISD) as a positive control for activation of TBK1 signaling lanes 1 and 2. (E) HDF, HDF-IFIT3, and HDF-IFIT3+JAK1 knockout cells were infected with HAdV-C5 at 200 P/cell and viral DNA replication was quantified by qPCR at 48 hpi, as described for Fig. 1C. The data are plotted as mean ± sd, n = 3. (**, P ≤ 0.01). (F) HDF, HDF-IFIT3, and HDF-IFIT3+JAK1 knockout cells were infected with HAdV-C5 at 200 P/cell and protein expression analyzed by Western blotting at 48 hpi using antibodies against total STAT1, phopsho-STAT1, JAK1, IFIT3, IFIT1, and Ad5 E1A.

Fig 5: JAK1 is not related to ISRE promoter activity in senescent cells. a ISRE activities are higher in NHDFs at passage 23 (p23) compared with passage 6 (p6). ISRE-driven luciferase expression constructs were transiently transfected, and a dual luciferase assay was performed. Data are expressed as means ± s.e. from two independent experiments performed in triplicate. *p < 0.05. b JAK1 knockdown did not affect ISRE activity in NHDFs at passage 24, but STAT1 or STAT2 knockdown decreased the activity of ISRE. Data are means ± s.e. from two independent experiments performed in triplicate. *p < 0.05. c−e JAK1 knockdown did not affect the Mx1 protein level (c), but STAT1 or STAT2 knockdown decreased the Mx1 protein level (d, e). Representative results from three independent experiments are shown. f The ISG mRNA level was reduced by STAT1 or STAT2 knockdown but not by JAK1 knockdown in NHDFs at passage 24. IL6 was included as an unrelated gene. Data are expressed as the means ± s.e. from two independent experiments performed in triplicate. *p < 0.05