Fig. 8

Involvement of NF-κB pathways in dCas9-E-induced EDA expression. The cells transfected with dCas9-E and incubated with doxycycline (Dox; 2 μg/ml) for 48 h exhibited a significance difference in NF-κB downstream Sonic Hedgehog (Shh) and cyclin D1 mRNA (a) and protein (b) expression. Immunostaining was also performed to detect the activity of the NF-κB pathway. dCas9-E BM-MSCs induced more NF-κB translocated into the nuclei and then activated the expression of Shh and cyclin D1 (c). All data are normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and calibrated based on the BM-MSC group. ***p <0.001 for Dox-treated dCas9-E BM-MSCs versus BM-MSCs. Scale bar = 20 μm. DAPI 4’6-diamidino-2-phenylindole, sgRNA single-guide RNA