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

Journal: International immunopharmacology

Article Title: 3,6-Anhydro-L-galactose suppresses mouse lymphocyte proliferation by attenuating JAK-STAT growth factor signal transduction and G 1 -S cell cycle progression.

doi: 10.1016/j.intimp.2024.113998

Figure Lengend Snippet: Fig. 10. Kinetic analysis of the inhibitory effect of L-AHG on the JAK-STAT signaling in activated T and activated B cells. (A, C) Following stimulation of G0 T cells (1.5 × 106/mL) and G0 B cells (2.5 × 106/mL) by immobilized anti-CD3/anti-CD28 and immobilized anti-CD40 + soluble anti-IgM + IL-4, respectively, for the indicated time periods in the absence or presence of L-AHG, cells were harvested and subjected to western blot analysis of the status of the JAK-STAT pathway components including p-JAK1 (Y1034/1035), JAK1, p-STAT1 (Y701), STAT1, p-STAT3 (Y705), STAT3 and GAPDH in activated T cells and activated B cells. A representative result is shown; two additional experiments yielded similar results. (B, D) The statistical data of western blot analysis have been drawn as bar graphs based on relative intensity of each interesting protein band. * p < 0.05, ** p < 0.01, *** p < 0.005, compared with the control. # p < 0.05, compared with selected groups.

Article Snippet: The antip-CDK4 (T172) antibody was purchased from ABclonal Technology (Woburn, MA, USA) and anti-cyclin A2, anti-cyclin E2, and anti-STAT3 antibodies were purchased from Proteintech (Rosemont, IL, USA).

Techniques: Western Blot, Control

Journal: Nutrients

Article Title: Hydroxycitric Acid Inhibits Chronic Myelogenous Leukemia Growth through Activation of AMPK and mTOR Pathway

doi: 10.3390/nu14132669

Figure Lengend Snippet: Hydroxycitric acid promotes AMPK phosphorylation in CML cells. CML cell lines were treated for 24 h with different concentrations of HCA. An equal amount of protein from each condition was separated on SDS PAGE (12% gel) and an immunoblot was performed using specific antibodies against pT172 AMPK and total AMPK. Vinculin was used as an internal control. ( A ) K562 ( B ) MEG-01 ( C ) KYO-1 ( D ) SHK-1. For the K562, MEG-01, KYO-1, and SHK-1, samples were loaded in duplicate. One was probed with total AMPK and the another one was probed with pT172 AMPK. Upper vinculin is referred to as pAMPK, and lower vinculin is referred to as the total AMPK. The bar graph beside each figure panel reflects the band intensity evaluated as optical density and represented as fold change for treated vs. untreated cells normalized for vinculin. ** p < 0.01, * p < 0.05 treated vs. untreated cells.

Article Snippet: Endogenous AMPK was immunoprecipitated using AMPKα antibody (Cell-Signaling, Danvers, MA, USA; #2523S).

Techniques: Phospho-proteomics, SDS Page, Western Blot, Control

Journal: Nutrients

Article Title: Hydroxycitric Acid Inhibits Chronic Myelogenous Leukemia Growth through Activation of AMPK and mTOR Pathway

doi: 10.3390/nu14132669

Figure Lengend Snippet: AMPK interacts with ACLY. ( A ) Co-immunoprecipitation of AMPK and ACLY in K562 cells. Endogenous AMPK was immunoprecipitated with antibody against total AMPK followed by Western blotting with anti-AMPK and anti-ACLY (upper panel). Bar graph showing the ratio of optical density of immunoprecipitated (IP) band of ACLY and AMPK after normalization with the respective input (lower panel). ( B ) Co-immunoprecipitation of AMPK and ACLY in K562 cells upon treatment with HCA (upper panel). Bar graph of optical density of immunoprecipitated ACLY and AMPK in each condition normalized to respective input band (left lower panel). Ratio of optical density of immunoprecipitated (IP) band of ACLY and AMPK after normalization with the control (right lower panel).

Article Snippet: Endogenous AMPK was immunoprecipitated using AMPKα antibody (Cell-Signaling, Danvers, MA, USA; #2523S).

Techniques: Immunoprecipitation, Western Blot, Control

Journal: Nutrients

Article Title: Hydroxycitric Acid Inhibits Chronic Myelogenous Leukemia Growth through Activation of AMPK and mTOR Pathway

doi: 10.3390/nu14132669

Figure Lengend Snippet: HCA stimulates unfolded protein response pathway. Cell lysate was prepared from K562 cells treated with indicated concentration of HCA. Proteins were separated and immunoblotted with antibody against desired protein. ( A ) HCA induces concomitant activation of AMPK and mTORC1 pathways in K562 cells. Samples were loaded in duplicate. One of them was used to immunoblot with phosphor form and the other was used to immunoblot respective total protein. Upper vinculin is referred to as p-AMPK, p-p70S6K, and pS6 ribosomal protein; lower vinculin is referred to as total AMPK, p70S6K, and S6 ribosomal protein. ( B ) HCA-treated K562 show upregulation of unfolded protein response markers ATF4 and p-elF2α. Upper vinculin is referred to as peIF2α and ATF4; lower vinculin is referred to as eIF2α. The bar graph beside each figure panel reflects the band intensity evaluated as optical density and represented as fold change for treated vs. untreated cells normalized for vinculin. ** p < 0.01, * p < 0.05 treated vs. untreated cells.

Article Snippet: Endogenous AMPK was immunoprecipitated using AMPKα antibody (Cell-Signaling, Danvers, MA, USA; #2523S).

Techniques: Concentration Assay, Activation Assay, Western Blot

Journal: Naunyn-Schmiedeberg's archives of pharmacology

Article Title: Modulation of AMPK by esomeprazole and canagliflozin mitigates methotrexate-induced hepatotoxicity: involvement of MAPK/JNK/ERK, JAK1/STAT3, and PI3K/Akt signaling pathways.

doi: 10.1007/s00210-025-03908-3

Figure Lengend Snippet: Fig. 7 The regulation of the p-JAK1/p-STAT3 signaling pathway by ESOM and CANA. To verify the effect of ESOM and CANA on various groups, we measured A the relative protein expression of p-JAK1 via western blot analysis and B the relative protein expres- sion of p-STAT3 using western blot analysis. Data are shown as mean ± SE (n = 6). *, **, # indicate a significant difference between the control group, MTX group, and ESOM and CANA + MTX group, respec- tively, using one-way ANOVA test at a p value < 0.01 followed by Tukey

Article Snippet: The membranes were incubated at 4 °C overnight with 1:1000 dilutions of JAK1 antibody (Santa Cruz Biotechnology, sc-376996, USA), STAT3 antibody (Abcam Co., EPR787Y, UK), JNK antibody (Santa Cruz Biotechnology, sc-7345, USA), ERK1 antibody (Santa Cruz Biotechnology, sc-271270, USA), and p38 antibody (Biorbyt, orb127559, USA), respectively, then incubated at room temperature for 1 h with 1:2000 dilutions of HRP-linked anti-rabbit antibody (Cell Signaling Technology, 7074, USA).

Techniques: Expressing, Western Blot, Control