Journal: Frontiers in Pharmacology

Article Title: Berberine suppresses hepatocellular carcinoma progression by blocking IL-4-JAK1-STAT6-mediated M2 polarization of macrophage

doi: 10.3389/fphar.2025.1734201

Figure Lengend Snippet: Berberine inhibits the activation of the JAK-STAT6 signal pathway induced by H22 conditional medium. (A–F) Relative transcription levels of M2-type signature genes in BMDM, including Arg 1 (A) , Retnla (B) , Il10 (C) , Mrc1 (D) , Tgfb1 (E) , and Chil3 (F) . BMDM were treated with H22 conditional medium for 4 h. (G) Immunoblot analysis of Arg-1 and Retnla protein expression in BMDMs. (H,I) Quantitative analysis of (H) Arg-1 and (I) Retnla protein levels normalized to β-actin. (J) Immunoblot detection of p-STAT6 protein expression in BMDM. (K) Quantitative statistics of p-STAT6/STAT6 are shown. (L) Immunoblot detection of p-JAK1 protein expression in BMDM. (M) Quantitative statistics of p-JAK1/JAK1 are shown. Data (mean ± SEM) are representative of three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001 .

Article Snippet: Primary antibodies were incubated overnight at 4 °C, including: p-STAT6 (56554S, CST), t-STAT6 (9362S, CST), p-JAK1 (3331S, CST), t-JAK1 (3344S, CST), β-actin (8457S, CST), Arg-1 (89872SF, CST), Retnla (Abcam, AB39626).

Techniques: Activation Assay, Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Berberine suppresses hepatocellular carcinoma progression by blocking IL-4-JAK1-STAT6-mediated M2 polarization of macrophage

doi: 10.3389/fphar.2025.1734201

Figure Lengend Snippet: Berberine inhibits the activation of the JAK-STAT6 signal pathway induced by IL-4. (A–F) Relative transcription levels of M2-type signature genes in BMDMs stimulated by IL-4 (20 ng/mL) for 4 h. Significant changes were observed in Arg1 (A) , Retnla (B), Il10 (C) , Mrc1 (D) , Tgfb1 (E) , and Chil3 (F) . (G,H) Western blotting analysis of p-STAT6 protein expression in BMDMs. Representative immunoblots (G) and quantitative analysis of p-STAT6/STAT6 (H) are shown. (I,J) Content analysis of p-JAK1 protein expression in BMDMs. Representative immunoblots (I) and quantitative analysis of p-JAK1/JAK1 (J) are shown. (K) Co-immunoprecipitation analysis of JAK1-STAT6 interaction in BMDMs under IL-4 (20 ng/mL) stimulation with or without BBR (30 μM) treatment. Data (mean ± SEM) are representative of three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001 .

Article Snippet: Primary antibodies were incubated overnight at 4 °C, including: p-STAT6 (56554S, CST), t-STAT6 (9362S, CST), p-JAK1 (3331S, CST), t-JAK1 (3344S, CST), β-actin (8457S, CST), Arg-1 (89872SF, CST), Retnla (Abcam, AB39626).

Techniques: Activation Assay, Western Blot, Expressing, Immunoprecipitation

Journal: Frontiers in Pharmacology

Article Title: Berberine suppresses hepatocellular carcinoma progression by blocking IL-4-JAK1-STAT6-mediated M2 polarization of macrophage

doi: 10.3389/fphar.2025.1734201

Figure Lengend Snippet: Synergistic therapeutic effects of Berberine and Anti-PDL1 antibody in treating H22 tumor. (A) Schema of the mouse tumor model: Female Bal/bc mice were engrafted with H22 tumor cells (5 × 10 5 ), received intragastric (i.g.) BBR (10 mg/kg), intraperitoneal (i.p.) Anti-PD-L1 antibody (10 mg/kg), or combination treatment. Tumors were removed and analyzed on day 21. (B) Tumor volume was measured every 2 days from day 9 after tumor implantation (n = 6). (C,D) On day 21 after tumor cell implantation, the tumors in mice were removed and weighed. (n = 6). Tumor growth curves (C) and tumor weight (D) are shown. (E–L) Flow cytometry analysis for the percentages of CD45 + lymphocytes in tumor tissues (E) , Percentage of CD3 + cells within the gated CD45 + cells in melanoma tissues (F) , CD3 + CD4 + T cells (G) , CD3 + CD8 + T cells (H) , CD45 + CD11b + Ly6G − Ly6C + M-MDSC (I) , CD45 + CD11b + Ly6G − LY6C − F4/80 + TAMs (J) , CD11b + F4/80 + MHC-Ⅱ + macrophages (K) , and CD11b + F4/80 + CD206 + macrophages (L) , within CD45 + population in tumors. Data (mean ± SEM) are representative of three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001. Schematic diagram illustrating the role of BBR in inhibiting tumor progression. BBR suppresses M2 polarization of tumor-associated macrophages in the tumor microenvironment and enhances the tumor-killing capacity of cytotoxic T lymphocytes, thereby inhibiting tumor progression. Mechanistically, BBR binds to the JAK1 protein in macrophages, inhibiting IL-4-induced phosphorylation of the JAK1-STAT6 signaling axis. This inhibition reduces the secretion of alternative activation genes by macrophages and blocks their pro-tumorigenic functions.

Article Snippet: Primary antibodies were incubated overnight at 4 °C, including: p-STAT6 (56554S, CST), t-STAT6 (9362S, CST), p-JAK1 (3331S, CST), t-JAK1 (3344S, CST), β-actin (8457S, CST), Arg-1 (89872SF, CST), Retnla (Abcam, AB39626).

Techniques: Tumor Implantation, Flow Cytometry, Phospho-proteomics, Inhibition, Activation Assay

Journal: Cosmetics

Article Title: Broussonetia papyrifera Promotes Hair Growth Through the Regulation of β-Catenin and STAT6 Target Proteins: A Phototrichogram Analysis of Clinical Samples

doi: 10.3390/cosmetics7020040

Figure Lengend Snippet: Figure 2. Luciferase activity of reporter gene by B. papyrifera. (a) For TCF/LEF reporter gene assay, cells were treated with B. papyrifera (1–40 µg/ml) for 24. The positive control group was treated with minoxidil (0.4 µM) or tofactinib (0.4 µM); (b) IL-4 induced STAT6; and (c) IL-6 induced STAT3 reporter gene assay as determined by luciferase activity. Tofactinib (0.4 µM), a JAKs inhibitor, was treated as a positive control. Each assay is representative for 3 experiments. The asterisk(s) indicate a significant statistical significance (** p < 0.05).

Article Snippet: Polyclonal antibodies against total β-catenin, phospho-specific β-catenin (Thr41/Ser45), STAT6, and phosphor-specific STAT6 (Tyr641) were purchased from Cell Signaling Technology (Beverly, MA, USA) and β-actin antibody, minoxidil, and tofactinb were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Luciferase, Activity Assay, Reporter Gene Assay, Positive Control

Journal: Cosmetics

Article Title: Broussonetia papyrifera Promotes Hair Growth Through the Regulation of β-Catenin and STAT6 Target Proteins: A Phototrichogram Analysis of Clinical Samples

doi: 10.3390/cosmetics7020040

Figure Lengend Snippet: Figure 3. Effect of B. papyrifera on the level or activity of proteins implicated in hair growth. (a) The level of p-β-catenin and β-catenin was detected by Western blotting using specific antibodies in hHF DP cells; (b) The level of p-STAT6 and STAT6 was detected by Western blotting using specific antibodies in IL-4-induced HDPCs; (c) The level of p-STAT3 and STAT3 was detected by Western blotting using specific antibodies in IL-6-induced HDPCs. β-Actin protein was used as an internal control. Immunoblot assay was performed as described in the Materials and Methods. Each blot is representative for 3 experiments.

Article Snippet: Polyclonal antibodies against total β-catenin, phospho-specific β-catenin (Thr41/Ser45), STAT6, and phosphor-specific STAT6 (Tyr641) were purchased from Cell Signaling Technology (Beverly, MA, USA) and β-actin antibody, minoxidil, and tofactinb were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Activity Assay, Western Blot, Control

Journal: PLoS ONE

Article Title: NK-4 exerts selective regulatory effects on the activation and function of allergy-related Th2 cells

doi: 10.1371/journal.pone.0199666

Figure Lengend Snippet: NHDF were grown in confluent monolayer cultures in 12-well plates and were stimulated with 10 ng/ml IL-4 and 5 ng/ml TNF-α in the presence or absence of varying concentrations of NK-4 for 15 min. Phosphorylation of STAT6 was determined by immunoblotting whole-cell lysates using specific antibodies against the phosphorylated or total STAT6 protein. A representative blot is shown (A). The optical density ratio of phospho-STAT6 to total STAT6 is shown (B). Data from three independent experiments were combined and expressed as the means ± SD. ** p < 0.01 compared with control cultures. Original uncropped and unadjusted Western blots were provided as supplementary files ( and ).

Article Snippet: Membranes were probed with a 1:1000 dilution of anti-phospho-STAT6 (Tyr641) rabbit mAb (C11A12; Cell Signaling Technology, Danvers, MA).

Techniques: Phospho-proteomics, Western Blot, Control