Figure 6
From: SMC3 knockdown triggers genomic instability and p53-dependent apoptosis in human and zebrafish cells
Effect of Smc3 knockdown on the phosphorylation in vivo of histone H3. Embryos at 1-2-two cell stage were injected with the vehicle alone or with 8 ng/embryo smc3-MO and analyzed at 24 hpf. Embryos were fixed overnight in 4% para-formaldehyde and permeabilized by immersion in -20°C acetone for 10 min. After overnight incubation at 4°C with 1 μg/ml rabbit anti-pH3 antibody, the embryos were placed in 1 μg/ml HRP-conjugated goat anti-rabbit IgG solution and the immunocomplexes detected by reaction with DAB reagent. The punctuate staining pattern visible in the embryo tails identifies the localization of the pH3 antigen and thus of the mitotic cells.