Figure 2. Inducible YTHDC1 deletion in satellite cells (SCs) abolishes acute injury-induced muscle regeneration.
(A) Breeding scheme for generating YTHDC1-inducible knockout (iKO) and control (Ctrl) mice. (B) Schematic outline of the tamoxifen (TMX) administration used in the study and experimental design for testing the effect of YTHDC1 deletion on barium chloride (BaCl2)-induced muscle regeneration process. (C) Left: western blotting (WB) showing the deletion of YTHDC1 in ASC-48 hr from iKO but not Ctrl mice. Right: no obvious morphological difference was detected in iKO vs. Ctrl mice. (D) H&E staining of the above injured muscles at 0, 5, and 7 days post injury (dpi). Scale bar = 100 μm. (E) Left: immunostaining of eMyHC (red) and laminin (green) of the above injured tibialis anterior (TA) muscles at 5 and 7 dpi. Scale bar = 100 μm. Right: quantification of eMyHC-positive fibers per field. n = 3 mice per group. (F) Immunostaining of Pax7 (red) and laminin (green) on TA muscle sections at 0, 5, and 7 dpi. Scale bar = 100 μm. (G) Quantification of Pax7-positive SCs per field at 0, 5, and 7 dpi. n = 4 mice per group for 0 dpi, n = 3 mice per group for 5 and 7 dpi. Bars represent mean ± SD for all graphs. Statistical significance was determined using a two-tailed Student’s t-test.
Figure 2—figure supplement 1. Analyses of regeneration at 14 and 28 days post injury.
