Figure 4.

MNV-induced apoptosis in the presence of 25-HC. Replicate wells of RAW264.7 cells were infected with MNV at an MOI of (A) 0.05 or (B) 5 TCID₅₀/cell and immediately treated with 25-HC (6.6 µM). As controls, the replicate wells were infected with MNV without the addition of 25-HC or treated with 25-HC alone. To quantify apoptosis, the caspase 3/7 assay reagent was added directly to the wells immediately after infection and the caspase activity was measured by fluorescence at regular intervals using IncuCyte ZOOM FLR. Data are shown as mean caspase-3/7 positive cells per well and statistical analysis of MNV with 25-HC by comparison to the MNV-only sample (n = 4 ± SEM, * = p < 0.05, ** = p < 0.01).