Goat anti s100a10

Sections from the control group were permeabilized with PBS containing 0.1% Triton X-100 (PBST) and 2% fetal bovine serum and incubated overnight at room temperature with primary antibodies to netrin-G1 (1:500 goat anti-Netrin-G1a, R&D Systems) or p11 (1:100 goat anti-S100A10, R&D Systems) and 5-HT (1:1000 rabbit anti-5HT, Sigma) or p11 and anti-phospho-histone H3 (PH3) (1:200 rabbit anti-phospho S10, Abcam). Slides were then washed in PBST before applying secondary antibodies. Incubation with secondary antibodies (1:800 anti-goat HRP and 1:800 anti-rabbit Rhodamine Red, Jackson ImmunoResearch Laboratories) occurred at room temperature for 2 h in the dark. Slides were again washed in PBST prior to amplification. For amplification, slides were incubated in Tyramide Signal Amplification-Fluorescein solution (1:50; PerkinElmer, NEL701A001 KT) for 10 min at room temperature in the dark. A final series of washes with PBST was performed and the slides were mounted using Prolong Gold with DAPI (to visualize nuclei; Life Technologies, P36931).