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. 2018 Jun 8;7:e26039. doi: 10.7554/eLife.26039

Figure 7. Compensatory effect of SOXF factors in satellite cells on ex vivo culture and in vivo injury-induced regeneration.

(A–C) Quantification of transduced satellite cells with SOXF-encoding retroviruses after 72 hr in culture on EDL isolated myofibers. Adult control satellite cells were transduced with the eGFP-encoding retrovirus (CTRL-RV) and Sox17-knockout cells with CTRL-RV or SOXF-FL. Quiescence (A; PAX7), activation (B; MYOD), and differentiation (C; MYOG) were measured. In red, CTRL vs. KO comparison; in black, KO transduced with CTRL-RV vs. KO transduced with SOXF-FL. n ≥ 30 fibers/EDL per condition; ≥1000 satellite cells/EDL. CTRL, Sox17GFP/fl; KO, Pax3Cre/+;Sox17GFP/fl. (D) Schematic outline of the experimental procedure for electroporation into regenerating TA muscle of wild type mice. CTX, cardiotoxin; d, days. (E) Histology characterization by Hematoxylin and eosin (cell infiltration, top panel), Oil red O (fat infiltration, middle panel), and Sirius red (fibrosis, bottom panel) staining of cryosections from electroporated wild type adult TA muscles five days after injury. TA muscles were electroporated with control (CTRL, left) or dominant negative SOX17 construct (SOX17ΔCt, right). Insets show enlarged images of the indicated regions. Quantification of fat infiltration (Oil red O) and fibrosis (Sirius red) are indicated as proportion of stained area. Scale bars, 100 μm. (F) RT-qPCR analysis seven days after CTX injection. n ≥ 3 mice (≥ 5 different areas). Data expressed as mean ± s.e.m., statistically analyzed with Student’s unpaired t-test: ns, not significant; *, p<0.05; **, p<0.01; ***, p<0.001, compared to CTRL-RV in CTRL (red asterisks in A-C), CTRL-RV in KO (black asterisks in A-C) or CTRL (E).

Figure 7.

Figure 7—figure supplement 1. Muscle electroporation during injury-induced muscle regeneration.

Figure 7—figure supplement 1.

(A) Schematic outline of the experimental procedure. (B–C) Representative images of electroporated wild type adult TA muscles five (B) and ten (C) days (d) after cardiotoxin (CTX) injection. TA muscles were injected with either pCIG (CTRL; green) or pCIG-expressing a dominant negative SOXF construct (SOX17ΔCt; green) (Figure 7), together with TdTomato (red) to identify the electroporated area. Scale bars, 5 mm. (D) Representative images of electroporated muscle cryosections five (left) and ten (right) days after CTX injection. BF, brightfield. n = 3 mice.