Journal: Journal of enzyme inhibition and medicinal chemistry

Article Title: Dual inhibition of EGFR and IL-6-STAT3 signalling by miR-146b: a potential targeted therapy for epithelial ovarian cancer.

doi: 10.1080/14756366.2021.1963240

Figure Lengend Snippet: Figure 2. The expression patterns of EGFR and IL-6-STAT3 in ovarian cancer. (A) Analysis of of IL-6 expression in ovarian cancer samples from TCGA database at http://gepia.cancer-pku.cn/. (B) Analysis of STAT3 expression in ovarian cancer tissues from the https://www.oncomine.org database. (C, D) The mRNA and protein lev- els of EGFR in ovarian cancer cell lines as determined by qPCR and Western blotting. (E) Analyses of mRNA levels of IL-6 in ovarian cancer samples (n ¼ 24) and control samples (n ¼ 24). (F) The relative mRNA levels of IL-6 in ovarian cancer cell lines. (G) The protein level of IL-6 as determined by Western blotting in ovarian cancer cell lines. (H) The concentration of IL-6 in cell supernatants cell supernatants as determined by flow cytometry in ovarian cancer cell lines. (I) The relative mRNA levels of STAT3 in ovarian cancer cell lines. (J) The relative expression of p-STAT3 (Y705, S727) and STAT3 in ovarian cancer cell lines was examined by Western blotting, and GAPDH was used as the internal control. The data are expressed as the means ± SDs; ns: not significant; p < 0.05; p < 0.01; p < 0.001.

Article Snippet: The antibodies used for Western blotting were as follows: anti-GAPDH (#60004–1-Ig, Proteintech, Wuhan, China), anti-IL-6 (#66146-Ig, Proteintech, Wuhan, China), anti-STAT3 (#4904 CST, Danvers, MA, USA), anti-P-STAT3 (Y705) (#9145, CST, Danvers, MA, USA), anti-P-STAT3 (S727) (#49081, CST, Danvers, MA, USA), anti-EGFR (#4267, CST, Danvers, MA, USA), anti-Flag (F1804, Sigma, USA), HRP-conjugated anti-mouse IgG (#70-GAM007, MultiSciences, China), and horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (#70-GAR007, MultiSciences, China).

Techniques: Expressing, Western Blot, Control, Concentration Assay, Flow Cytometry

Journal: Theranostics

Article Title: IL20RA signaling enhances stemness and promotes the formation of an immunosuppressive microenvironment in breast cancer.

doi: 10.7150/thno.45280

Figure Lengend Snippet: Figure 4. IL20RA promotes breast cancer stemness via STAT3 activation. (A-B) Western blot results of p-STAT3(Tyr705), STAT3, p-JAK1, JAK1, OCT4, SOX2 and IL20RA in T-47D cells treated with STAT3 inhibitor (CDDO-Im) or JAK1 inhibitor (filgotinib) for 48 h. (C) Statistical analysis of the proportion of SP in T-47D cells treated with CDDO-Im and filgotinib, respectively (n = 3 for each group). (D) Statistical analysis of the sphere-formation in T-47D cells, which were treated with CDDO-Im and filgotinib, respectively (n = 3 for each group). (E-H) BALB/c mice bearing 4T1-IL20RA allografts were treated with DMSO and CDDO-Im, respectively. (E) Tumor growth curves (n = 6 for each group). (F) Upper panel: primary tumors separated from each group at the end of experiments. Lower panel: statistical analysis of tumor weights. Data are presented as mean ± SEM (n = 6 for each group). (G) Left panel: representative IHC staining images of p-STAT3 (Tyr705) and SOX2 in 4T1-IL20RA allograft tissues. Scale bar, 20 µm. Right panel: statistical analyses of the IHC-scores of p-STAT3 (Tyr705) and SOX2 (n=6 for each group) in paraffin-embedded tumor tissue sections. (H) Left panel: representative H&E staining images of lung metastasis in 4T1-IL20RA allograft mice, scale bar, 100 µm. Right panel: statistical analysis of the lung metastatic foci number (n = 6 for each group). *, p < 0.05; **, p < 0.01.

Article Snippet: IHC staining was performed on tumor tissues of mice using antibodies against IL20RA, p-STAT3 (Tyr705) and CD8α (Santa Cruz Biotechnology, Dallas, TX, USA), SOX2 (Proteintech, Wuhan, Hubei, China) and CD11c (Cell Signaling Technology, Theranostics 2021, Vol.

Techniques: Activation Assay, Western Blot, Immunohistochemistry, Staining

Journal: Theranostics

Article Title: IL20RA signaling enhances stemness and promotes the formation of an immunosuppressive microenvironment in breast cancer.

doi: 10.7150/thno.45280

Figure Lengend Snippet: Figure 6. Combination of immunotherapy and targeted therapy with chemotherapy for breast cancer with high IL20RA expression. (A) Schematic diagram of the experimental procedure. (B-D) BALB/c mice bearing 4T1-IL20RA allografts were treated with DOX plus CTX combined with IgG, αPD-L1 Ab, NPs-Stattic, NPs-Stattic-IL20RA plus IgG, and NPs-Stattic-IL20RA plus αPD-L1 Ab, respectively. (B) Tumor growth curves (n = 6 mice for each group). Tumor volumes between different groups are compared 32 days after inoculation to determine the statistical significances. (C) The image of primary tumors separated from different groups at the end of experiments. (D) Statistical analysis of tumor weights at the end of experiments. Data are presented as mean ± SEM, n = 6 mice for each group. (E) Body weight curves of BALB/c mice from different groups (n = 6 mice for each group). (F) Statistical analyses of the IHC-scores of p-STAT3 (Tyr705), SOX2, CD11c and CD8α in paraffin-embedded tumor tissue sections. Data are presented as mean ± SEM, n = 6 mice for each group. *, p < 0.05; **, p < 0.01; ns, not significant.

Article Snippet: IHC staining was performed on tumor tissues of mice using antibodies against IL20RA, p-STAT3 (Tyr705) and CD8α (Santa Cruz Biotechnology, Dallas, TX, USA), SOX2 (Proteintech, Wuhan, Hubei, China) and CD11c (Cell Signaling Technology, Theranostics 2021, Vol.

Techniques: Expressing

Journal: Cancer biology & therapy

Article Title: GdX inhibits the occurrence and progression of breast cancer by negatively modulating the activity of STAT3.

doi: 10.1080/15384047.2024.2420383

Figure Lengend Snippet: Figure 6. GdX specifically interacts with STAT3. (a) Dual luciferase assay results indicate that STAT3 is a downstream target gene of GdX; (b) immunofluorescence staining reveals the colocalization of GdX and p-STAT3 in the cell nucleus; (c) Western blot analysis assesses the impact of GdX overexpression on the phosphorylation levels of STAT3. *, p < .05, **, p < .01, ***, p < .001, vs. GdX-nc.

Article Snippet: The membrane was blocked with 5% skimmed milk for 1 h at room temperature, and then incubated with primary antibodies against GdX (Proteintech, 1: 1000), Bcl-XL (Proteintech, 1: 1000), c-Myc (Proteintech, 1: 2000), Cyclin D1 (Proteintech, 1: 5000), STAT3 (Cell Signaling Technology, 1: 1000), phospho-STAT3 (p-STAT3) (Cell Signaling Technology, 1: 2000), and GAPDH (Cell Signaling Technology, 1: 1000) at 4°C overnight.

Techniques: Luciferase, Immunofluorescence, Staining, Western Blot, Over Expression, Phospho-proteomics