Figure 6

Knockdown of CAV1 affects apoptosis of iPAM cells by ApxI protein. A CAV1 protein levels were assessed by siRNA silencing and western blotting. A single 50 nM target-specific siRNA was transfected into iPAM cells for 24 h. Lysates were separated by SDS‒PAGE and detected by western blotting with an antibody against CAV1. B Western blot detection of the activated caspase 3 band (17 kDa) with a caspase 3 antibody. The average intensity of active caspase 3 in the immunoblot was quantified and normalized to the intensity of β-actin. C The effect of CAV1 gene silencing on ApxI toxin-involved apoptosis was examined under fluorescence microscopy using the Annexin-V-FITC Apoptosis Assay Kit. iPAM cells transfected with or without CAV1 siRNA were exposed to 50 μg/mL ApxI toxin for 34 h. Graphs show the mean cell apoptosis for each group. A t test showed highly significant changes between the control and knockdown groups (p = 0.0013) (p < 0.01).