Anti mouse igg h l alexa fluor 647

Immunofluorescence was performed on OECs. The cells were fixed with 4% formalin solution at room temperature for 5 min then washed with PBS and permeabilized with 0.1% Triton X-100. The cells were then blocked with 1% bovine serum albumin (10082-139; Gibco; Thermo Fisher Scientific, Inc.) at 37°C for 30 min and incubated for 18 h at 4°C with anti-p75NGFR (1:500; AB1554; EMD Millipore) and anti-BrdU (1:200; mAb #5292; Cell Signaling Technology, Inc., Danvers, MA, USA) antibodies. Following washing with PBS, the cells were incubated at 37°C for 2 h with respective anti-Mouse IgG H&L (Alexa Fluor 647; 1:500; ab150115; Abcam, Cambridge, MA, USA). Following incubation, the slides were washed with PBS, mounted and examined under a fluorescence microscope (Leica TCS SP2; Leica Microsystems GmbH, Wetzlar, Germany). Fluoroshield mounting medium with DAPI (Sigma-Aldrich; Merck KGaA) was used for DAPI staining and the staining was performed according to the manufacturer's protocol.