Additional file 3 of Tetraspanin CD82 is necessary for muscle stem cell activation and supports dystrophic muscle function
figureposted on 2020-11-27, 04:51 authored by Arielle Hall, Tatiana Fontelonga, Alec Wright, Katlynn Bugda Gwilt, Jeffrey Widrick, Alessandra Pasut, Francesco Villa, Cynthia K. Miranti, Devin Gibbs, Evan Jiang, Hui Meng, Michael W. Lawlor, Emanuela Gussoni
Additional file 3: Supplementary Figure 3. Delayed fusion in CD82-/- myoblast cultures is not due to early apoptosis/or reduced cell adhesion. (A, B) Immunofluorescence staining of cultures for MyoD (red) and H2Aγ (green) 72 hrs post-extraction from WT (A) and CD82-/- (B) animals. Arrowhead in (A) points at a MyoD+H2Aγ+ cell. (C) Quantification of H2Aγ+ cells and (D) non-adherent cells revealed no significant differences between the genotypes at all timepoints analyzed. (E) Myogenic cells from WT and (F) CD82-/- mice were plated at the same density and induced to form myotubes. At day 3 and 5 following differentiation, WT and CD82-/- cultures were immunostained for myosin heavy chain (red staining in G, H, L, M) to quantify the fusion index. The purity of the cultures assessed by MyoD staining was >80-90%. At day 3 there was no difference in myotube formation between cultures, as assessed by fusion index following staining with MF20 (I). (N) Quantification of fusion index after 5 days in differentiation media showed significantly decreased fusion in CD82-/- compared to WT cultures. Fusion index was calculated as the ratio of number of nuclei fused in MHC-myotubes over the number of total nuclei (****p<0.0001).