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

Journal: Nutrients

Article Title: Dietary Soy Protein Isolate Attenuates Intestinal Immunoglobulin and Mucin Expression in Young Mice Compared with Casein

doi: 10.3390/nu12092739

Figure Lengend Snippet: Primers for qRT-PCR.

Article Snippet: The following antibodies were used: anti-Janus kinase-1 (JAK1) (bs-1439R, Bioss, Beijing, China), anti-phospho (p)-JAK1 (Tyr1034 + Tyr1035, bs-3238R, Bioss, China), anti- signal transducer and activator of transcription 6 (STAT6) (380957, ZEN BIO, China), anti-p-STAT6 (Tyr641, ab263947, Abcam, Cambridge, MA, USA), anti-pIgR (AF2800-SP, R&D Systems, Minneapolis, MN, USA), anti-TGFβ1 (bs-4538R, Bioss, China), and anti-β-actin (AP0060, Bioworld Technology Inc., Bloomington, MN, USA).

Techniques:

Journal: Nutrients

Article Title: Dietary Soy Protein Isolate Attenuates Intestinal Immunoglobulin and Mucin Expression in Young Mice Compared with Casein

doi: 10.3390/nu12092739

Figure Lengend Snippet: Effect of SPI on JAK1-STAT6 signaling and TGFβ levels in mouse ileum tissue. ( A ) qRT-PCR analysis of Jak1 , Stat6 , Tgfb1 , and Tgfb2 mRNA expression levels in mouse ileum tissue (normalized against Actb (β-actin); n = 8). ( B ) Western blotting analysis of JAK1, p -JAK1, STAT6, p -STAT6, pIgR, and TGFβ1 protein levels in mouse ileum tissue ( n = 6). ( C ) The statistical analyses of the Western blotting results. Results are normalized to β-actin ( n = 6). All data are presented as means ± SEM. NS, not significant ( p > 0.05); * p < 0.05; ** p < 0.01. Statistical significance was calculated using t -tests.

Article Snippet: The following antibodies were used: anti-Janus kinase-1 (JAK1) (bs-1439R, Bioss, Beijing, China), anti-phospho (p)-JAK1 (Tyr1034 + Tyr1035, bs-3238R, Bioss, China), anti- signal transducer and activator of transcription 6 (STAT6) (380957, ZEN BIO, China), anti-p-STAT6 (Tyr641, ab263947, Abcam, Cambridge, MA, USA), anti-pIgR (AF2800-SP, R&D Systems, Minneapolis, MN, USA), anti-TGFβ1 (bs-4538R, Bioss, China), and anti-β-actin (AP0060, Bioworld Technology Inc., Bloomington, MN, USA).

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: Scientific Reports

Article Title: 3-oxo-C12:2-HSL, quorum sensing molecule from human intestinal microbiota, inhibits pro-inflammatory pathways in immune cells via bitter taste receptors

doi: 10.1038/s41598-022-13451-3

Figure Lengend Snippet: 3-oxo-C12:2-HSL prevents activation of the JAK-STAT pathway. Cells were stimulated with LPS and IFNγ in absence (control) or in presence of 50 µM 3-oxo-C12:2-HSL for 2 h. The levels of phosphorylated proteins P-JAK1 ( a ), STAT1 ( b ), JAK2 ( c ), STAT3 ( d ) were normalized to their respective unphosphorylated forms. Results are expressed as mean ± SEM from 4 independent experiments. One-Way ANOVA, Dunnett’s post-test **p < 0.01, ***p < 0.001, ****p < 0.0001 vs. control. ( e ) Reconstructed images from Simple Western analysis of protein levels and actin used as housekeeping protein are displayed. They are based on the area under the chemiluminescence signal curve obtained for one experiment, representative of 4 independent experiments performed in duplicates. Molecular markers are indicated on the left (kDa). 3oC12:2 stands for 3-oxo-C12:2-HSL.

Article Snippet: The microplates were loaded with 0.8 μg/μL protein, primary antibodies (as detailed below), and reagents provided by the manufacturer: anti-JAK1 (1:25; Biotechne MAB4260), anti-Phospho-JAK1 (1:100; Invitrogen 44422G), anti-JAK2 (1:13; Cell Signalling, 3230), anti-Phospho-JAK2 (1:13; Cell Signalling 3776), anti-STAT1 (1:50; Cell Signalling 9172), anti-Phospho-STAT1 (1:40; Biotechne AF2894), anti-Actin (1:1000; Millipore MAB1501R).

Techniques: Activation Assay, Western Blot

Journal: Scientific Reports

Article Title: 3-oxo-C12:2-HSL, quorum sensing molecule from human intestinal microbiota, inhibits pro-inflammatory pathways in immune cells via bitter taste receptors

doi: 10.1038/s41598-022-13451-3

Figure Lengend Snippet: Proposed mechanisms of 3-oxo-C12:2-HSL effects on inflammation. In immune cells, 3-oxo-C12:2-HSL is able to activate bitter taste receptors, which are G-protein coupled receptors, triggering a signalling cascade resulting in the release of calcium from the endoplasmic reticulum. In addition, in murine activated macrophages the presence of 3-oxo-C12:2-HSL attenuates the activation of the JAK-STAT signalling pathways, by specifically preventing JAK1 and STAT1 phosphorylation. This leads to a decrease in pro-inflammatory cytokine secretions and an overall reduced inflammatory response. Part of the effects of 3-oxo-C12:2-HSL on inflammatory response is dependent on bitter taste receptors. Created with BioRender.com.

Article Snippet: The microplates were loaded with 0.8 μg/μL protein, primary antibodies (as detailed below), and reagents provided by the manufacturer: anti-JAK1 (1:25; Biotechne MAB4260), anti-Phospho-JAK1 (1:100; Invitrogen 44422G), anti-JAK2 (1:13; Cell Signalling, 3230), anti-Phospho-JAK2 (1:13; Cell Signalling 3776), anti-STAT1 (1:50; Cell Signalling 9172), anti-Phospho-STAT1 (1:40; Biotechne AF2894), anti-Actin (1:1000; Millipore MAB1501R).

Techniques: Activation Assay