Fig. 3

dCas9-E mediated activation of EDA in BM-MSCs. The cells were harvested and total mRNA and protein were extracted. Ectodysplasin (EDA) gene and protein expression were identified by qPCR (a) and Western blotting (b) after doxycycline (Dox) induction for 48 h. c Fluorescence microscopy detection of EDA expression activated by dCas9-E. After supplementing with 2 μg/ml Dox for 48 h, enhanced expression of EDA (red) was detected in dCas9-E transfected bone marrow-derived mesenchymal stem cells (BM-MSCs). The data are expressed as the mean ± SD. ***p < 0.001 for Dox-treated cells versus untreated cells by Dunnett’s test. Scale bar = 50 μm. DAPI 4’6-diamidino-2-phenylindole, GAPDH glyceraldehyde-3-phosphate dehydrogenase, sgRNA single-guide RNA