Fig. 5

Wnt3a-induced Ins2 upregulation is blocked by knockdown of NeuroD1. NeuroD1 was knocked down by infecting N39 cells with lentivirus containing shRNA for 72 h. a N39 cells were infected by lentiviral shRNA against NeuroD1 for 48 h and then treated with Wnt3a (100 ng/mL) for 24 h. The levels of active and total β-catenin and NeuroD1 protein were determined by immunoblot assay. b The intensity of NeuroD1 signal in (a) was quantified using the ImageJ software (n = 4). The levels of NeuroD1 mRNA (c) and Ins2 mRNA (d) were measured by qRT-PCR in N39 cells that were infected with lentiviral non-targeting or NeuroD1 shRNA for 48 h and then treated with Wnt3a (100 ng/mL) for 24 h (n = 9). Data are means + SEM. ***p < 0.001 compared with vehicle treatment in shNon-target. ###p < 0.001 compared with Wnt3a treatment in shNon-target. N.S., not significant within shNeuroD1 samples