NLRP3 Inflammasome: Key Mediator of Neuroinflammation in Murine Japanese Encephalitis (original) (raw)
Figure 6
Generation of ROS is critical for caspase-1 activity and subsequent IL-1β and IL-18 maturation.
BV-2 cells were incubated with 1 µM of DPI for inhibition of ROS generation. (A) Representative FACS plot showing intracellular ROS production, after 4 h of JEV infection and representation of the Mean Fluorescent Intensities (M.F.I) (right graph) in mock-infected control (C), JEV+DPI and JEV alone condition. (B) Caspase-1 activity measured after 6 h of JEV infection in presence or absence of DPI. Graph represents fold change in caspase-1 activity with respect to mock-infected control. (C–D) ELISA study showing the levels of mature IL-1β and IL-18 in JEV+DPI condition with respect to JEV alone infected sample. Graph represents cytokine levels in pg/ml. For Potassium efflux study, BV-2 cells were incubated with 50 mM KCl for 20 min in order to study the requirement of K**+** efflux for caspase-1 activity and its downstream effects. Data represent mean ± SEM from 3 independent experiments performed in duplicate. Statistical differences were evaluated using one way ANOVA with Bonferroni's post hoc test. *, **, Statistical difference in comparison to mock-infected control values (*p<0.05, ** p<0.01) and #, Statistical difference with respect to JEV infected condition (p<0.01).