pax7 ab 528428 myod mouse igg2b  (Developmental Studies Hybridoma Bank)

Journal: Skeletal Muscle

Article Title: TRPV1 manipulating polarization of M1/M2 macrophages to promote skeletal muscle regeneration

doi: 10.1186/s13395-026-00417-6

Figure Lengend Snippet: Activation of TRPV1 facilitates myogenesis during the process of muscle regeneration in vivo. A-C Representative western blot and relative protein level of MyoD and myogenin in CTX-induced muscle tissue after treatment with CAP and CPZ at different points in time ( n = 3 animals per experimental group; mean ± SD; Two-way ANOVA). D , F The expression level of MyoD and myogenin mRNA in each group at different points in time ( n = 3 animals per experimental group; mean ± SD; Two-way ANOVA). E Representative immunofluorescence images of MyoD in CTX-induced muscle tissue after treatment with CAP and CPZ at 4d. G-H The relative fluorescence intensity of MyoD and the proportion of MyoD+ /DAPI + double-positive cells in each group ( n = 5 animals per experimental group; mean ± SD; One-way ANOVA). Statistical significance was set at P < 0.05. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Scale bar, 50 μm

Article Snippet: Deparaffinized sections were blocked with 5% bovine serum albumin (BSA) and incubated overnight at 4 °C with the following primary antibodies: mouse anti-CD86 monoclonal antibody (1:200, Santa Cruz Biotechnology, sc-28347); mouse anti-CD206 polyclonal antibody (1:200, Santa Cruz Biotechnology, sc-58986); rat anti-F4/80 monoclonal antibody (1:100, Abcam, ab1691); rabbit anti-F4/80 monoclonal antibody (1:200, Proteintech, #27044-1-AP); rabbit anti-TRPV1 polyclonal antibody (1:200, ABclonal, A8564); and mouse anti-MyoD monoclonal antibody (1:100, Santa Cruz Biotechnology, sc-377460).

Techniques: Activation Assay, In Vivo, Western Blot, Expressing, Immunofluorescence, Fluorescence

Journal: Skeletal Muscle

Article Title: TRPV1 manipulating polarization of M1/M2 macrophages to promote skeletal muscle regeneration

doi: 10.1186/s13395-026-00417-6

Figure Lengend Snippet: The influence of TRPV1 in the C2C12 myoblasts differentiation. A-C The relative fluorescence intensity of TRPV1( n = 5 independent random fields of cells; mean ± SD; One-way ANOVA). D Representative western blot of TRPV1 after treatment with CAP and CPZ in vitro. E The protein expression of TRPV1 ( n = 3 independent replicates in cells; mean ± SD; One-way ANOVA). F Representative western blot of MyoD and myogenin after treatment with CAP and CPZ in differentiation medium at different intervals. G , H The protein expression of MyoD and myogenin among each group ( n = 3 independent replicates in cells; mean ± SD; Two-way ANOVA). I , J The relative fluorescence intensity of MYH3 in multinucleated myotubes in each group time-dependently ( n = 5 independent random fields of cells; mean ± SD; Two-way ANOVA). K Representative Giemsa staining images showed the effect of TRPV1 activation individually on multinucleated myotubes ( n = 5 independent random fields of cells). Statistical significance was set at P < 0.05. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Scale bar, 50 μm, 100 μm

Article Snippet: Deparaffinized sections were blocked with 5% bovine serum albumin (BSA) and incubated overnight at 4 °C with the following primary antibodies: mouse anti-CD86 monoclonal antibody (1:200, Santa Cruz Biotechnology, sc-28347); mouse anti-CD206 polyclonal antibody (1:200, Santa Cruz Biotechnology, sc-58986); rat anti-F4/80 monoclonal antibody (1:100, Abcam, ab1691); rabbit anti-F4/80 monoclonal antibody (1:200, Proteintech, #27044-1-AP); rabbit anti-TRPV1 polyclonal antibody (1:200, ABclonal, A8564); and mouse anti-MyoD monoclonal antibody (1:100, Santa Cruz Biotechnology, sc-377460).

Techniques: Fluorescence, Western Blot, In Vitro, Expressing, Staining, Activation Assay

Journal: Skeletal Muscle

Article Title: TRPV1 manipulating polarization of M1/M2 macrophages to promote skeletal muscle regeneration

doi: 10.1186/s13395-026-00417-6

Figure Lengend Snippet: TRPV1 regulates M1/M2 macrophage polarization to promote myogenic differentiation in C2C12 cells. A C2C12 myoblasts were co-cultured with M1 or M2 macrophages for 4 days via a transwell cell culture insert. B Representative Western blot bands of MyoD and MYH3 in C2C12 myoblasts after being co-cultured with M1 or M2 macrophages for 4 days. C , D The protein expression of MyoD and MYH3 in those C2C12 myoblasts which were co-cultured with M1or M2 macrophages after CAP and CPZ treatment ( n = 3 independent replicates in cells; mean ± SD; Two-way ANOVA). E , F Representative immunofluorescence images showed myotubes fusion index in C2C12 cells ( n = 5 independent random fields of cells; mean ± SD; Two-way ANOVA). Statistical significance was set at P < 0.05. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Scale bar, 50 μm

Article Snippet: Deparaffinized sections were blocked with 5% bovine serum albumin (BSA) and incubated overnight at 4 °C with the following primary antibodies: mouse anti-CD86 monoclonal antibody (1:200, Santa Cruz Biotechnology, sc-28347); mouse anti-CD206 polyclonal antibody (1:200, Santa Cruz Biotechnology, sc-58986); rat anti-F4/80 monoclonal antibody (1:100, Abcam, ab1691); rabbit anti-F4/80 monoclonal antibody (1:200, Proteintech, #27044-1-AP); rabbit anti-TRPV1 polyclonal antibody (1:200, ABclonal, A8564); and mouse anti-MyoD monoclonal antibody (1:100, Santa Cruz Biotechnology, sc-377460).

Techniques: Cell Characterization, Cell Culture, Western Blot, Expressing, Immunofluorescence

Journal: Journal of Advanced Research

Article Title: Versatile platforms of mussel-inspired agarose scaffold for cell cultured meat

doi: 10.1016/j.jare.2025.01.024

Figure Lengend Snippet: Proliferation of C2C12 cells on AG/PDA gel scaffold. a) Representative images of C2C12 cells stained with EdU incorporation assay on AG/PDA gel scaffolds, b) EdU positive cells of C2C12 cells were evaluated (n = 6). The PDA coating has optical absorption properties, color codes: blue: nuclei; red: EdU, multicolor: merge, bars: 100 μm. c) Western blot analysis of MYOD and GADPH protein levels in the proliferative and differentiation phases (n = 3). d ) Quantification of MYOD expression normalized to GAPDH from western blot gel (n = 3). Data presented as mean ± SD (* p < 0.05, ** p < 0.01 and *** p < 0.001, ns: not significant).

Article Snippet: After sealing the membranes with 5 % bovine serum albumin buffer for 2 h at room temperature, the membranes were incubated at 4 °C overnight with primary antibodies ((rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Abcam, ab181602, 1:10000), mouse anti- myogenic differentiation antigen (MyoD) (Santa Cruz Biotechnology, sc-377460, 1:500), primary MYHC antibody (1:500), then secondary antibodies (horseradish peroxidase (HRP)-conjugated goat anti-rabbit (Beyotime, A208,1:1000), HRP-conjugated goat anti-mouse antibody (Beyotime, A216, 1:1000) was incubated for 1 h at room temperature.

Techniques: Staining, Western Blot, Expressing

Journal: Journal of Advanced Research

Article Title: Versatile platforms of mussel-inspired agarose scaffold for cell cultured meat

doi: 10.1016/j.jare.2025.01.024

Figure Lengend Snippet: Myogenesis of PSMSCs on AG/PDA gel scaffold. a) Representative photographic images of PMSCs were first adhesion after inoculation on AG/PDA gel scaffolds for two hours. b) Adhesion ratio of PSMSCs on AG/PDA gel scaffolds (n = 6). c) Representative images of PSMSCs stained with EdU incorporation assay on AG/PDA gel scaffolds. d) EdU positive cells of PSMSCs were evaluated (n = 6). The PDA coating has optical absorption properties, color codes: blue: nuclei; red: EdU, multicolor: merge, bars: 50 μm. e) the metabolic activity of PSMSCs with alma blue cell viability assay on AG/PDA gel scaffolds (n = 6). f) Representative images of immunofluorescent stained PSMSCs and myotubes after 5 days of differentiation on AG/PDA gel scaffolds. Color codes: blue: nuclei, red: MYHC, green: phalloidin, multicolor: merge, bars: 50 μm. g) Western blot analysis of MYHC, MYOD, and GAPDH protein levels in the proliferative and differentiation phases (n = 3). h) Quantification of MYHC and MYOD expression normalized to GAPDH from western blot gel (n = 3). Data presented as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001, ns: not significant).

Article Snippet: After sealing the membranes with 5 % bovine serum albumin buffer for 2 h at room temperature, the membranes were incubated at 4 °C overnight with primary antibodies ((rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Abcam, ab181602, 1:10000), mouse anti- myogenic differentiation antigen (MyoD) (Santa Cruz Biotechnology, sc-377460, 1:500), primary MYHC antibody (1:500), then secondary antibodies (horseradish peroxidase (HRP)-conjugated goat anti-rabbit (Beyotime, A208,1:1000), HRP-conjugated goat anti-mouse antibody (Beyotime, A216, 1:1000) was incubated for 1 h at room temperature.

Techniques: Staining, Activity Assay, Viability Assay, Western Blot, Expressing

Journal: Journal of Advanced Research

Article Title: Versatile platforms of mussel-inspired agarose scaffold for cell cultured meat

doi: 10.1016/j.jare.2025.01.024

Figure Lengend Snippet: Myogenesis of PSMSCs on AG/PDA film scaffold. a) Schematic of the production process of film scaffolds. b) Representative SEM images of the PDA particles on film scaffold, bar: 1 μm. c) Analysis of the surface hydrophilic angle on film scaffolds (n = 3). d) Representative images of surface hydrophilic angles of film scaffolds and Water absorption swelling rate of film scaffolds (n = 3). e) Representative SEM images of the AG/PDA-0.2 film scaffold surface with and without PSMSCs adhesion, bar: 1 μm. f) the metabolic activity of PSMSCs with alma blue cell viability assay on AG/PDA film scaffolds (n = 6). g) Representative 3D reconstruction images of immunofluorescent stained PSMSCs and myotubes after 5 days differentiation on AG/PDA film scaffolds. Color codes: blue: nuclei, red: MYHC, green: phalloidin, multicolor: merge, bars: 150 μm. h) Western blot analysis of MYHC, MYOD, and GAPDH protein levels in the proliferative and differentiation phases (n = 3). i) Quantification of MYHC and MYOD expression normalized to GAPDH from western blot gel (n = 3). Data presented as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001, ns: not significant).

Article Snippet: After sealing the membranes with 5 % bovine serum albumin buffer for 2 h at room temperature, the membranes were incubated at 4 °C overnight with primary antibodies ((rabbit anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Abcam, ab181602, 1:10000), mouse anti- myogenic differentiation antigen (MyoD) (Santa Cruz Biotechnology, sc-377460, 1:500), primary MYHC antibody (1:500), then secondary antibodies (horseradish peroxidase (HRP)-conjugated goat anti-rabbit (Beyotime, A208,1:1000), HRP-conjugated goat anti-mouse antibody (Beyotime, A216, 1:1000) was incubated for 1 h at room temperature.

Techniques: Activity Assay, Viability Assay, Staining, Western Blot, Expressing