Fig. 2

ASD-associated de novo R1008X and Q1042R mutations disrupt the DNA-binding activity of POGZ. a POGZ is localized to the nucleus in dissociated embryonic cortical neurons (7 days in vitro). left, Double immunostaining for POGZ (green) and a neuron marker, Tuj1 (red). Nuclei were stained with Hoechst 33258 (blue). Scale bar, 10 μm. right, Biochemical preparation of the cytosolic and nuclear fractions from dissociated neurons. Equal quantities of protein were loaded into individual lanes and probed with antibodies against POGZ, GAPDH (a cytosolic marker), and Histone H3 (a nuclear marker). b Wild-type (WT) but not R1008X POGZ binds a CENP-B box sequence. left, Representative western blots. right, Quantification of DNA-binding activity. Notably, the Q1042R mutation led to a reduction of approximately 60 % in DNA binding (each n = 6, *p < 0.05, **p < 0.01, one-way ANOVA followed by Tukey-Kramer post hoc tests (vs. WT)). The averaged WT value was set to 100 %. Data are expressed as the means ± SEM