Fig. 2

Knocking-out Yy1 reduced proliferation of MHB NECs. a, b Immunofluorescent staining detecting 1-h labelled BrdU at E10.5. c Statistical analysis showing the percentage of BrdU-positive cells is reduced in mutant E10.5 MHB neuroepithelium. Two-tailed unpaired t-test, ***, p = 0.002. At least two sections counted from 4 pairs of 4 litters. d, e Staining of phospho-Histone H3 and Tuj1. f Cumulative BrdU-labeling experiments at E10.5 showing the lengthened cell-cycle time in mutants (control Tc = 720.1 min; mutant Tc = 910.1 min; correlation coefficients r2(control) = 0.9814, and r2(mutant) = 0.8861). g Representative flow cytometry results of control and mutant MHB dissociated cells stained with PI and analyzed by ModFit LT. Blue arrowhead pointing to the debris in mutant samples. h Statistical analysis of MHB cell cycle. Control, n = 10; Mutant, n = 7. Two-tailed unpaired t-test, *, p = 0.039; ***, p < 0.001. i Significant upregulated genes relative mRNA expression level. N numbers are shown in the bar of controls. Unpaired t-test, ***, p < 0.001. j, k Immunofluorescent imaging showing P21 expression increased in the mutant mes/r1 region