Antibody stains were performed as previously described51 (link) and the pigment on fish was bleached after the antigen retrieval step with a 1% H2O2/3% KOH solution. Antibodies used were total ERK (1:500, Cell Signaling 4696) and phospho-histone H3 (1/500, Millipore 06-570).
For western blotting, whole adult brains were dissected and homogenized in RIPA buffer with protease inhibitors. Anti-α-tubulin from Sigma (T6074-200UL (020M4753)) was used as the control at 1/40,000 for approximately 45 min at room temperature, followed by horseradish-peroxidase-coupled mouse secondary antibody for 1 h at room temperature. RhoA antibody from Cytoskeleton (http://www.cytoskeleton.com/arh04 ) was used at 1/500 overnight at 4 °C. Immunohistochemistry was performed as in ref. 51 (link) with RhoA antibody from Cytoskeleton (http://www.cytoskeleton.com/arh04 ) freshly diluted in PBS at 1/100.
For western blotting, whole adult brains were dissected and homogenized in RIPA buffer with protease inhibitors. Anti-α-tubulin from Sigma (T6074-200UL (020M4753)) was used as the control at 1/40,000 for approximately 45 min at room temperature, followed by horseradish-peroxidase-coupled mouse secondary antibody for 1 h at room temperature. RhoA antibody from Cytoskeleton (