2.11 Confocal assay
LX‐2 cells were plated on glass cover slips in a dish. After exposure to
different treatments, the cells were washed three times with PBS, fixed
with 4% paraformaldehyde for 20 min and then permeabilized with 0.5%
Triton X‐100 for 30 min at room temperature. Next, the cells were
incubated with specific primary antibodies overnight at 4°C. Cells were
washed three times with PBS and subsequently incubated with a
fluorochrome‐labelled secondary antibody for 1 hr at room temperature.
The cells were then rinsed three times with PBS and the nuclei were
counterstained with DAPI for 5 min at room temperature. Images were
captured using a Leica SP2 confocal microscope (Leica Microsystems,
Exton, PA)