Journal: Advanced Science
Article Title: Cilia‐Mediated Insulin/Akt and ST2/JNK Signaling Pathways Regulate the Recovery of Muscle Injury
doi: 10.1002/advs.202202632
Figure Lengend Snippet: IL13 promotes muscle regeneration and M2 polarization. IL13 or PBS was intraperitoneally injected into glycerol‐injected WT mice at 1 dpi. A) Schematic of the experimental schedule. B) Immunofluorescence analysis of CD68 + M1 (M1 MΦ, red) or CD206 + M2 (M2 MΦ, red), laminin (green), and nuclei (Hoechst33342, blue) in TA muscle cross sections at 3 dpi. C,D) Quantification of the CD68‐stained area ( n = 5 or 6) (C) or the CD206‐stained area ( n = 8) (D) per 40× view in (B). TA muscle cross‐sections of PBS‐ or IL13‐injected mice at 3 dpi. Scale bars, 40 µm. E,F) Quantification of average cross‐section area of myofiber in TA muscle of PBS‐ ( n = 5) or IL13‐injected mice ( n = 8) at 3 dpi (E) and representative images of immunofluorescence for laminin (green) and nuclei (Hoechst33342, blue) (F). Scale bars, 400 µm (left) and 20 µm (right). G–I) Immunofluorescence analysis of CD68 + M1 (M1 MΦ, red) or CD206 + M2 (M2 MΦ, red), laminin (green), and nuclei (Hoechst33342, blue) in TA muscle cross sections of glycerol‐injected WT and Tchp −/− mice at 3 dpi (G) and quantification of CD68‐stained area ( n = 5 or 6) (H) or CD206‐stained area ( n = 8) (I) per 40× view. Scale bar: 50 µm. All data are the mean ± S.D. * p < 0.05, ** p < 0.01, N.S., not significant; two‐tailed unpaired Student's t ‐tests.
Article Snippet: Staining was performed using the following antibodies: acetylated tubulin (6‐11B‐1, Sigma–Aldrich; 1:200), Arl13b (17711‐1‐AP, Proteintech; 1:200), PTRF (18892‐1‐AP, Proteintech; 1:100), Caveolin‐1 (#3238, Cell Signaling Technology; 1:300), Collagen I (ab21286, Abcam; 1:100), Perilipin (D1D8, Cell Signaling Technology; 1:200), CD45 and CD45‐biotin (30‐F11, Biolegend; 1:100), CD68 (FA‐11, Bio‐Rad; 1:100), CD206 (MR5D3, Bio‐Rad; 1:100), Desmin (D93F5, Cell Signaling Technology; 1:200), Flotillin‐2 (B‐6, Santa Cruz Biotechnology; 1:20), FOXP3 (FJK‐16s, Invitrogen; 1:100), GM3 (GMR6, Tokyo Chemical Industry; 1:500), IL4 (BVD4‐1D11, Novus Biologicals; 1:100), IL13 (AF‐413, R&D Systems; 1:100), IL33 (AF3626, R&D Systems; 1:100), PAX7 (MAB1675, R&D Systems; 1:100), PDGFR α (AF1062, R&D Systems; 1:200; APA5, Biolegend; 1:100), p‐Histone H3 (Ser 10) (sc‐8656‐R, Santa Cruz Biotechnology; 1:100), Sca1 (E13‐161.7, Biolegend, 1:100), Vimentin (#280618, R&D Systems; 1:200), Ki67 (SolA15, Invitrogen; 1:200), and Laminin (L9393, Sigma Aldrich; 1:200).
Techniques: Injection, Immunofluorescence, Staining, Two Tailed Test
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