Fig. 2: NANOG expression de-differentiates skeletal muscle to a pro-regenerative state.

The left leg of naïve uninjured mice was implanted with Dox-Elvax for two weeks; the right leg was implanted with DMSO-Elvax and served as control for the Elvax placement. One week after removal of Elvax, the TA muscle was harvested for evaluation. A Immunostaining of TA muscle sections for eMYHC (red), Laminin (green) and DAPI (Blue). Scale bar = 100 µm, insets are higher magnification images with scale bar = 50 µm. B Quantification of fiber area of TA muscle in mice with Elvax-DOX vs. Elvax-DMSO; n = 2 biological replicates for both DMSO-Elvax and Dox-Elvax, 200-250 muscle fibers from 3 muscle sections. C Quantification of small muscle fibers (less than 100 µm²) expressing eMYHC; n = 7 fields of view for 2 biological replicates for both DMSO-Elvax and Dox-Elvax. D Percentage of centrally nucleated regenerative muscle fibers throughout the TA muscle; n = 2 biological replicates for both DMSO-Elvax and Dox-Elvax, 200-250 muscle fibers per mouse. E Immunostaining for Pax7 (red), Laminin (green) and DAPI (blue). Fields of view depicted are representative images from the center of the muscle, and section adjacent to Elvax placement. Scale bar = 20 µm. F Quantification for percentage of Pax7 positive nuclei throughout the TA muscle; n = 2 biological replicates for both DMSO-Elvax and Dox-Elvax. G Western blot for atrogenes Atrogin-1 and Murf-2B, Murf-2A and Murf-1/3 in the TA muscle. Fold change in (H) Atrogin-1 (I) Murf-2B (J) Murf-2A and (K) Murf-1/3 expression in Dox-Elvax compared to DMSO-Elvax quantified after normalization to GAPDH; n = 4 biological replicates for both DMSO-Elvax and Dox-Elvax. Unpaired t-test used for statistical analysis in all graphs. All data are presented as Mean ± SEM. Source data are provided as a Source Data file.