emyhc  (Developmental Studies Hybridoma Bank)

Journal: The Journal of Clinical Investigation

Article Title: Activin A secretion by muscle-repairing macrophages induces heterotopic ossification in mice

doi: 10.1172/JCI193797

Figure Lengend Snippet: ( A ) Experimental timeline of the transfer experiment. DT (400 ng/mouse) was administered intraperitoneally on the indicated days to LysM-Cre iDTR mice to induce macrophage depletion. Ly6C hi CX3CR1 lo PDPN + CD9 + MDMs and other MDMs (the remaining MDM population) were isolated from muscle tissues of WT mice at 1 dpi. The isolated cells, suspended in 10 μL PBS or the same volume of PBS alone, were then injected intramuscularly into the recipient mice. ( B and C ) Representative images of H&E ( B ) and immunofluorescence staining ( C ) of muscle sections from DT-treated LysM-Cre iDTR mice injected intramuscularly with PBS ( n = 3), other MDMs ( n = 5), or Ly6C hi CX3CR1 lo PDPN + CD9 + MDMs ( n = 6) at 4 dpi. Muscle sections were stained with anti-mouse eMyHC (red) to mark regenerating muscle fibers and Hoechst (blue) to label nuclei. The boxed areas in the main images are shown at higher magnification (×4) in the adjacent lower panels. H&E staining and corresponding immunofluorescence images represent the same field of view. Scale bars: 100 μm. ( D and E ) Quantification of muscle regeneration showing the distribution of the CSA of eMyHC + fiber ( D ) and the mean CSA of eMyHC + fibers in an individual mouse ( E ). The P values were calculated using 1-way ANOVA with Tukey’s multiple-comparison test. A P value < 0.05 was considered significant. Data are shown as the mean ± SEM, and symbols represent individual fibers ( D ) or individual mice ( E ).

Article Snippet: For the evaluation of muscle regeneration, sections were stained with anti-mouse embryonic myosin heavy chain (eMyHC) (catalog 22287-1-AP; Proteintech) and corresponding Alexa Fluor 594 secondary antibody (catalog A-11012; Thermo Fisher Scientific) to visualize regenerating fibers.

Techniques: Isolation, Injection, Immunofluorescence, Staining, Comparison

Journal: The Journal of Clinical Investigation

Article Title: Activin A secretion by muscle-repairing macrophages induces heterotopic ossification in mice

doi: 10.1172/JCI193797

Figure Lengend Snippet: ( A and B ) Representative images of H&E and immunofluorescence staining of muscle sections from Inhba fl/fl mice ( n = 8) and Inhba fl/fl LysM-Cre mice ( n = 8) at 4 dpi. In immunofluorescence staining, sections were stained for anti-mouse eMyHC (red) to indicate regenerating fibers and Hoechst (blue) to indicate nuclei. The boxed areas are magnified (×4) in the panels to the right. H&E staining and corresponding immunofluorescence images represent the same field of view. Scale bars: 100 μm. ( C – E ) Quantification of regeneration showing the distribution of the CSA of eMyHC + fiber ( C ), the mean CSA of eMyHC + fibers in an individual mouse ( D ), and the percentage of regeneration area (calculated as total eMyHC + area/injured area) ( E ). ( F and G ) Representative images of H&E and immunofluorescence staining of muscle sections from Inhba fl/fl mice ( n = 8) and Inhba fl/fl LysM-Cre mice ( n = 8) at 4 dpi. In immunofluorescence staining, sections were stained for mIgG (white) to indicate necrotic fibers. H&E staining and corresponding immunofluorescence images represent the same field of view. Scale bars: 50 μm. ( H ) The density of mIgG + fibers (calculated as the number mIgG + fibers/injured area). ( I ) Grip strength of the forelimb in Inhba fl/fl and LysM-Cre Inhba fl/fl mice at 7 dpi. gf, gram force. ( J and K ) Photographs of gastrocnemius of the Inhba fl/fl ( J ) and LysM-Cre Inhba fl/fl ( K ) mice at 14 dpi. Scale bars: 5 mm. ( L ) Mass of gastrocnemius of Inhba fl/fl and LysM-Cre Inhba fl/fl mice at 14 dpi. The P values were calculated using unpaired 2-tailed t test. A P value < 0.05 was considered significant. Data are shown as the mean ± SEM, and symbols represent individual fibers ( C ) or individual mice ( D , E , H , I , and L ).

Article Snippet: For the evaluation of muscle regeneration, sections were stained with anti-mouse embryonic myosin heavy chain (eMyHC) (catalog 22287-1-AP; Proteintech) and corresponding Alexa Fluor 594 secondary antibody (catalog A-11012; Thermo Fisher Scientific) to visualize regenerating fibers.

Techniques: Immunofluorescence, Staining

Journal: The Journal of Clinical Investigation

Article Title: Activin A secretion by muscle-repairing macrophages induces heterotopic ossification in mice

doi: 10.1172/JCI193797

Figure Lengend Snippet: ( A – C ) H&E and immunofluorescence staining of muscle sections from WT ( A ; n = 4), Tlr4 –/– ( B ; n = 5), and Ticam1 –/– mice ( C ; n = 5) at 4 dpi. Immunofluorescence staining showing eMyHC-labeled (red) regenerating muscle fibers and Hoechst-stained (blue) nuclei. The boxed areas are shown at higher magnification (×4) in the panels to the right. H&E staining and the corresponding immunofluorescence images represent the same field. Scale bars: 100 μm. ( D and E ) Quantification of regeneration showing the distribution of eMyHC + CSA ( D ) and the mean CSA of eMyHC + fibers ( E ). The P values were calculated using 1-way ANOVA with Tukey’s multiple-comparison test. A P value < 0.05 was considered significant. Data are shown as the mean ± SEM, and symbols represent individual fibers ( D ) or individual mice ( E ).

Article Snippet: For the evaluation of muscle regeneration, sections were stained with anti-mouse embryonic myosin heavy chain (eMyHC) (catalog 22287-1-AP; Proteintech) and corresponding Alexa Fluor 594 secondary antibody (catalog A-11012; Thermo Fisher Scientific) to visualize regenerating fibers.

Techniques: Immunofluorescence, Staining, Labeling, Comparison