Immunofluorescence Analysis of Apoptosis and Cell Markers

For immunofluorescence double staining, apoptosis was detected by the ApopTag Fluorescein In Situ Apoptosis Detection Kit (S7110; Chemicon International, Temecula, CA), as described previously^{15 (link)}. And then, sections were incubated overnight with cell surface glycoprotein F4/80 (Serotec, Oxford, UK) and a Texas red-labeled secondary antibody and counterstained with 4,6-diamidino-2-phenylindole (DAPI). We also performed immunofluorescence double staining for SOX10 (Abcam, Cambridge, UK), β3-tubulin (1:50; Cell Signaling Technology, Danvers, MA), and LC3 (1:200; Sigma-Aldrich, St. Louis, MO, USA). The fluorescent images were examined under a laser scanning confocal microscope system (Carl Zeiss LSM 700, Oberkochen, Germany).