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: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Sanguinarine mediated apoptosis in Non-Small Cell Lung Cancer via generation of reactive oxygen species and suppression of JAK/STAT pathway.

doi: 10.1016/j.biopha.2021.112358

Figure Lengend Snippet: Fig. 4. SNG downregulates constitutively activated JAK/STAT3 signaling pathway and IL6 mediated STAT3 activation in NCI-H-1975 and HCC-827 cellS (A). After 24 h of SNG treatment, NCI-H-1975 and HCC-827 cells were lysed, and western blotting was performed for antibodies against p-Jak2, Jak2, GAPDH, p-Src, Src, p- STAT3 (Try705 and ser727), and STAT3. (B) siRNA knockdown of STAT3. NCI-H-1975 were transfected with control (100 pM) and STAT3 siRNA (50 and 100 pM). Cells were lysed and separated, transferred on the PVDF membrane, and immunoblotted with antibodies against STAT3, Bcl-xL, caspase-3, cleaved caspase-3 and HSP60. (C) SNG inhibits IL 6 secretion in NCI-H-1975 and HCC-827 cells. The cells mentioned above were exposed to SNG for 24 h, as indicated. Using supernatant from the treated and untreated group, levels of IL 6 were determined using multiplex biometric ELISA-based immunoassay. (D) SNG inhibits IL6-induced STAT3 activation. NCI-H-1975 and HCC-827 cells were pretreated with 2.0 μM SNG for 1 h and then stimulated with IL6 (100 µg/ml). At the end of the treatment, cells were lysed and immunoblotted against p-STAT3 (Try705) and STAT3(ser727).

Article Snippet: Antibodies against caspase-9 (Cat. No. 9508), Bcl2 (Cat. No. 15071), phospho-STAT3 Tyr705- Cat. No. 9145, phospho-STAT3 Ser727- Cat. No. 9134, STAT3 Cat. No.12640, cleaved caspase-3 Cat. No. 9661, caspase-3 Cat. No. 9662, Bax (Cat. No.5023), Cytochrome c (Cat. No.12963), Tubulin (Cat. No.2144), GAPDH (Cat. No.5174), PARP ( Cat. No.9542) CLEAVED CASPASE 8 (Cat. No.9496), phospho-jak2 (Cat. No.3776), JAK2(Cat. No.3230), PH2AX(Cat. No.9718), phsopho-Src Tyr416 (Cat. No. 6943) and Src (Cat. No. 2108) were purchased from Cell Signaling Technologies (Beverly, USA).

Techniques: Activation Assay, Western Blot, Knockdown, Transfection, Control, Membrane, Multiplex Assay, Enzyme-linked Immunosorbent Assay

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Sanguinarine mediated apoptosis in Non-Small Cell Lung Cancer via generation of reactive oxygen species and suppression of JAK/STAT pathway.

doi: 10.1016/j.biopha.2021.112358

Figure Lengend Snippet: Fig. 7. SNG inhibits NCI-H-1975 xenograft tumor growth in ICR-SCID mice. Subcutaneous tumor was grown in donor mouse and then transplanted into ICR-SCID female mice and divided into treated and control groups as dis cussed under Materials and Methods. At the end of the study, the mice were sacrificed, and (A) excised tumors in each group were weighed and analyzed using graph pad prism software. The graph displays the mean ± SD of four inde pendent experiments (* P < 0.05). (B) Excised tumors were photographed (C) lysates obtained from excised tumors were immunoblotted against p-STAT3, STAT3, and actin antibodies. (D) Densitometric analysis of p-STAT3/STAT3 expression in control and SNG treated tumors. The graph displays the mean ± SD of four in dependent experiments (* P < 0.05).

Article Snippet: Antibodies against caspase-9 (Cat. No. 9508), Bcl2 (Cat. No. 15071), phospho-STAT3 Tyr705- Cat. No. 9145, phospho-STAT3 Ser727- Cat. No. 9134, STAT3 Cat. No.12640, cleaved caspase-3 Cat. No. 9661, caspase-3 Cat. No. 9662, Bax (Cat. No.5023), Cytochrome c (Cat. No.12963), Tubulin (Cat. No.2144), GAPDH (Cat. No.5174), PARP ( Cat. No.9542) CLEAVED CASPASE 8 (Cat. No.9496), phospho-jak2 (Cat. No.3776), JAK2(Cat. No.3230), PH2AX(Cat. No.9718), phsopho-Src Tyr416 (Cat. No. 6943) and Src (Cat. No. 2108) were purchased from Cell Signaling Technologies (Beverly, USA).

Techniques: Control, Software, Expressing

Journal: International journal of oral science

Article Title: Parathyroid hormone increases alveolar bone homoeostasis during orthodontic tooth movement in rats with periodontitis via crosstalk between STAT3 and β-catenin.

doi: 10.1038/s41368-020-00104-2

Figure Lengend Snippet: Fig. 3 Daily systemic injection of PTH activated STAT3 in the alveolar bone, which could be reversed by local stattic injection. Representative images of immunohistochemical staining of STAT3 (a) and p-STAT3 (Tyr705) (b) in the OTM + PD, OTM + PD + PTH and OTM + PD + PTH + S groups, and relative quantitative analysis at days 7 and 14. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.000 1 by one-way ANOVA with Tukey’s post hoc test

Article Snippet: The membranes were blocked with 5% skim milk or 5% BSA and incubated with primary antibodies against β-actin (Huabio, China), STAT3 (Huabio, China), phospho-STAT3 (Tyr705) (Cell Signaling Technology, USA), RUNX2 (Huabio, China), OSX (Abcam, UK), ALP (Huabio, China), OPN (Huabio, China), β-catenin (Huabio, China), RANKL (Abcam, UK) and OPG (Huabio, China) at 4 °C overnight, and rabbit and mouse secondary antibodies (Signalway Antibody, USA).

Techniques: Injection, Immunohistochemical staining, Staining

Journal: International journal of oral science

Article Title: Parathyroid hormone increases alveolar bone homoeostasis during orthodontic tooth movement in rats with periodontitis via crosstalk between STAT3 and β-catenin.

doi: 10.1038/s41368-020-00104-2

Figure Lengend Snippet: Fig. 6 IPTH promoted osteogenesis in vitro, which could be reduced by stattic. a Alp, Ibsp, Opn, Sp7, Pth1r, Cbfa1, Col1a1 and Bglap expression in each group. b Protein levels of RANKL and OPG. c RANKL and OPG mRNA levels and the RANKL/OPG ratio. d Protein levels of representative osteoblastic markers and p-STAT3 (Tyr705) and STAT3. e Alizarin red S staining and ALP staining. f Quantitative analysis of Alizarin red S staining. g Quantitative analysis of ALP. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.000 1 by one-way ANOVA with Tukey’s post hoc test

Article Snippet: The membranes were blocked with 5% skim milk or 5% BSA and incubated with primary antibodies against β-actin (Huabio, China), STAT3 (Huabio, China), phospho-STAT3 (Tyr705) (Cell Signaling Technology, USA), RUNX2 (Huabio, China), OSX (Abcam, UK), ALP (Huabio, China), OPN (Huabio, China), β-catenin (Huabio, China), RANKL (Abcam, UK) and OPG (Huabio, China) at 4 °C overnight, and rabbit and mouse secondary antibodies (Signalway Antibody, USA).

Techniques: In Vitro, Expressing, Staining

Journal: International journal of oral science

Article Title: Parathyroid hormone increases alveolar bone homoeostasis during orthodontic tooth movement in rats with periodontitis via crosstalk between STAT3 and β-catenin.

doi: 10.1038/s41368-020-00104-2

Figure Lengend Snippet: Fig. 7 IPTH activated Wnt/β-catenin signalling, while inhibition of STAT3 activation suppressed this effect. a Colocalization of β-catenin and STAT3 by immunofluorescence. b β-catenin levels in nuclear fractions (active form) and whole cells. c Axin2 expression level. d Ctnnb1 expression level. e Sost expression level. f A diagram showing the hypothesis that PTH regulates STAT3 and Wnt/β-catenin signalling. *P < 0.05, **P < 0.01 by one-way ANOVA with Tukey’s post hoc test

Article Snippet: The membranes were blocked with 5% skim milk or 5% BSA and incubated with primary antibodies against β-actin (Huabio, China), STAT3 (Huabio, China), phospho-STAT3 (Tyr705) (Cell Signaling Technology, USA), RUNX2 (Huabio, China), OSX (Abcam, UK), ALP (Huabio, China), OPN (Huabio, China), β-catenin (Huabio, China), RANKL (Abcam, UK) and OPG (Huabio, China) at 4 °C overnight, and rabbit and mouse secondary antibodies (Signalway Antibody, USA).

Techniques: Inhibition, Activation Assay, Expressing