Mouse anti vc1

Whole-mount immunohistochemistry staining was performed as previously described^{121 (link)}. Animals were sacrificed with 2% HCl and fixed with 4% FA. Animals were blocked in 1% bovine serum albumin (BSA) in 1X PBSTx 0,3% (Blocking Solution) for 2 h at RT. Primary antibodies were diluted in blocking solution and incubated 16 h rocking at 4 °C. Then, washes were per performed for at least 4 h. Secondary antibodies were diluted in blocking solution for 16 h rocking at 4 °C.
The following antibodies were used in these experiments: mouse anti-synapsin (anti-SYNORF1, 1:50; Developmental Studies Hybridoma Bank, Iowa City, IA, USA), rabbit anti-phospho-histone H3 (Ser10) (D2C8) (PH3) (1:500; Cell Signaling Technology) and mouse anti-VC1 (anti-arrestin, 1:15,000, kindly provided by Professor K. Watanabe). The secondary antibody used was Alexa 488-conjugated goat anti-mouse (1:400; Molecular Probes, Waltham, MA, USA) and Alexa 568-conjugated goat anti-rabbit (1:1000: Molecular Probes, Waltham, MA, USA). Nuclei were stained with DAPI (1:5000; Sigma).

Whole-mount immunohistochemistry staining was carried out as previously described [35 (link)]. Animals were sacrificed with 2% HCl, fixed with 4% FA and blocked in 1% bovine serum albumin (BSA) in 1× PBST × 0.3% (blocking solution) for 2 h at RT. Primary antibodies were incubated in blocking solution for 16 h rocking at 4 °C. Washes were per performed for at least 4 h, and secondary antibodies were diluted in blocking solution for 16 h rocking at 4 °C. The following antibodies were used in these experiments: mouse anti-synapsin (anti-SYNORF1, 1:50; Developmental Studies Hybridoma Bank, Iowa City, IA, USA), mouse anti-VC1 (anti-arrestin, 1:15000, kindly provided by Professor K. Watanabe) and rabbit anti-phosphohistone H3 (Ser10) (D2C8) (PH3) (1:500; Cell Signaling Technology, Leiden, Netherlands). The secondary antibodies used were Alexa 488-conjugated goat anti-mouse (1:400; Molecular Probes, Waltham, MA, USA) and Alexa 568-conjugated goat anti-rabbit (1:1000: Molecular Probes, Waltham, MA, USA). Nuclei were stained with DAPI (1:5000; Sigma).