Journal: Frontiers in Veterinary Science

Article Title: γ -Aminobutyric acid enhances myogenesis and heat/cold stress resistance in bovine muscle satellite cells

doi: 10.3389/fvets.2026.1770540

Figure Lengend Snippet: Effects of GABA on the proliferative activity and molecular markers in BSCs. (A) Immunofluorescence staining for BSCs identification (Desmin, Pax7, and MyoG (green) and nuclei (DAPI, blue) (10×; scale bar = 200 μm). (B–E) CCK-8 analysis of viability after 24, 48, 72, and 96 h treatment with 0.1, 0.5, 1, 2, 5, and 10 mM GABA and CON = 0 mM. (F,G) EdU assay. Hoechst (nuclei, blue) and EdU-positive cells (red) scale bar = 200 μm. (H–M) RT-qPCR. mRNA expression of proliferation-related genes ( CDK2 , PCNA , Ki-67 , Pax7 , Pax3 , CCND1 ) after 48 h treatment with 0.5 and 1 mM GABA. (N,O) Western blotting of PAX7, CDK1, CDK2, and P21 proteins (β-actin loading control). (Mean ± SD; n = 3; groups with different letters a–e differs at p < 0.05).

Article Snippet: Polyclonal antibodies against myogenin (MYOG; BS-3550R, Bioss, Beijing, China), paired box protein 7 (Pax7; AB-528428, Abcam, United Kingdom) and desmin (BS-20702R, Bioss, Beijing, China) were each diluted 1:100 in PBS containing 5% bovine serum albumin (BSA) and incubated overnight at room temperature.

Techniques: Activity Assay, Immunofluorescence, Staining, CCK-8 Assay, EdU Assay, Quantitative RT-PCR, Expressing, Western Blot, Control

Journal: Cancer Medicine

Article Title: Transgelin Expression in Activated Cancer‐Associated Fibroblasts Regulates Stromal Contractility and Promotes Colon Cancer Progression

doi: 10.1002/cam4.71789

Figure Lengend Snippet: Transgelin expression pattern in colorectal cancer. (A) Transgelin expression in normal colonic tissues. (B) Transgelin expression in colorectal cancer tissues. (C) Transgelin expression in cancer tissues. (D) Fluorescent immunostaining showed that transgelin was expressed in desmin‐positive smooth muscles as well as in other cancer stromal regions; transgelin, desmin are shown in dark green, green, and purple, respectively. (E) Immunostaining results for transgelin (dark green), smooth muscle Actin (SMA; red), and 4′,6‐diamidino‐2‐phenylindole (DAPI; purple) in the cancer cell stroma. (F) Immunostaining for AE1/3 (green), transgelin (dark green), and DAPI (purple).

Article Snippet: The primary antibodies used were mouse monoclonal anti‐human transgelin antibody (OriGene Technologies, Rockville, MD, USA; used at 1:200 dilution), rabbit monoclonal anti–human α‐SMA (SPRING BIOSCIENCE, Pleasanton, CA, USA; used at 1:200 dilution), Desmin (Nichirei, Tokyo, Japan; used without dilution), 4′,6‐diamidino‐2‐phenylindole (DAPI; Akoya Biosciences, Menlo Park, CA, USA; used without dilution), and AE1/3 antibody (Dako, Agilent Technologies, Santa Clara, CA, USA; used without dilution).

Techniques: Expressing, Immunostaining, Muscles

Journal: Journal of Advanced Research

Article Title: Podocyte TLR4 deletion alleviates diabetic kidney disease through prohibiting PKCδ/SHP-1-dependent ER stress and relieving podocyte damage and inflammation

doi: 10.1016/j.jare.2025.07.013

Figure Lengend Snippet: Blockade of TLR4/MyD88/PKCδ/SHP-1 signaling attenuated podocyte cytoskeletal remodeling and inflammation, and macrophage recruitment and infiltration through decreasing ER stress and MCP-1 production under hyperglycemic conditions. (A) Assessment of podocyte migration by transwell assay in cultured podocytes treated with LG or HG for 48 h with or without transfection of TLR4, PKCδ, MCP-1 or scrambled shRNAs, or pretreatment of 4-PBA. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ## P < 0.01; ### P < 0.01; #### P < 0.01 ( n = 3 independent experiments). (B) Representative IF images by phalloidin staining to illustrate F-actin distribution and cytoskeletal remodeling in cultured podocytes treated with LG or HG for 48 h with or without transfection of TLR4, PKCδ, MCP-1 or scrambled shRNAs, or pretreatment of 4-PBA. Magnification 400 × . ( n = 3 independent experiments). (C) Assessment of the secretion of inflammatory factors IL-6 and IL-1β in podocytes treated with LG or HG for 48 h with or without transfection of TLR4, PKCδ, or scrambled shRNAs, or pretreatment of 4-PBA. P values were determined by one-way ANOVA followed by Tukey’s multiple comparisons test and data are presented as mean ± SD. **** P < 0.01; # P < 0.05; ## P < 0.01; ### P < 0.01 ( n = 3 independent experiments). (D) IHC staining using anti-f4/80 antibodies to illustrate macrophage infiltration in the glomeruli from mice at 16 weeks (Red arrow). Scale bar, 20 μm. ( n = 8) (E) The model diaphragm of podocyte-macrophage co-culture. (F) Representative images and assessment of macrophage migration by transwell assay after co-cultured with podocytes treated with LG or HG for 48 h with or without transfection of TLR4, PKCδ, MCP-1 or scrambled shRNAs, or pretreatment of 4-PBA. Magnification 400 × . P values were determined by one-way ANOVA followed by Tukey’s multiple comparisons test and data are presented as mean ± SD. **** P < 0.01; ## P < 0.01; ### P < 0.01 ( n = 3 independent experiments).

Article Snippet: The following primary antibodies were used: f4/80 (29414–1-AP), desmin (16520–1-AP), MCP-1 (26161–1-AP), podocin (20384–1-AP), synaptopodin (21064–1-AP), TLR4 (66350–1-Ig), β-actin (66009–1-Ig), PKCδ(14188–1-AP), ATF6 (24169–1-AP), p-cadherin (13773–1-AP) were from Proteintech (IL, USA); phospho-PKCδ (Y311) (ab76181), PKCdelta (ab182126), SHP-1 (ab227503), HA (ab9110) were from Abcam (MA, USA); BIP (M010300), MyD88 (P012343) were from Epizyme, Shanghai, China); and ATF4 (11815), sXBP-1 (40435) were from Cell Signaling Technology (MA, USA).

Techniques: Migration, Transwell Assay, Cell Culture, Transfection, Staining, Immunohistochemistry, Co-Culture Assay