2.6. Immunofluorescence

The hPDLSCs were seeded in a 12-well plate under different treatments. Cells were fixed and permeabilized using 4% paraformaldehyde and 0.1% Triton X-100. After blocking with 1% bovine serum albumin (BSA), the cells were incubated with cleaved Caspase-3 (c-Caspase-3) (Cell Signaling, USA) as primary antibody at room temperature for 4 h. Then, the cells were incubated with the secondary antibody conjugated to Alexa555 (Invitrogen, USA). Followed by incubation with Hoechst (Life Technologies, USA), the cells were imaged using fluorescence microscopy (Carl Zeiss).