Figure 3.

Cul3 modulates cell viability through caspase-3 activity in primary cultured hippocampal neurons. (A) Representative images of WT, heterozygous, and homozygous fCul3 cultured hippocampal neurons treated with Z-DEVD-FMK or vehicle transduced with AAV-Cre-GFP viruses (WT-Cre, Het-Cre, Hom-Cre). DAPI, cell nucleus marker. Cl. Caspase-3, cleaved caspase-3. Scale bar, 50 μm. (B) Neuron number counted based on DAPI signal. (C) AAV virus transduction efficiency was comparable among all groups. (D) Caspase-3 positive signal was divided by DAPI signal to determine the caspase-3-positive cell rate. Increased caspase-3 activity is largely reversed by Z-DEVD-FMK treatment in Hom group. (E) Representative images of WT, heterozygous, and homozygous fCul3 cultured hippocampal neurons treated with Z-DEVD-FMK or vehicle transduced with AAV-Cre-GFP viruses (WT-Cre, Het-Cre, Hom-Cre). Calcein, viable cell marker. Scale bar, 50 μm. (F) Neuron number based on DAPI signal. (G) AAV virus transduction efficiency was comparable among all groups. (H) Calcein positive signal was divided by DAPI signal to determine viable cell rate. Decreased viable cell rate is largely reversed by Z-DEVD-FMK treatment in both Het and Hom group. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ordinary two-way AVONA with Dunnett’s multiple comparisons test; graphs depict mean ± SEM. N = 30 neurons from 3 separately derived neuronal cultures (10 neurons for each group).