Figure 4.

HIRA knockdown promotes NPC terminal mitosis. (A) The timeline shows the process of the BrdU birthdating experiment. (B and C) Control or HIRA shRNA plasmids were electroporated into embryonic mouse brains at E13.5, and BrdU was injected at E14.5. Then, the electroporated embryonic brains were collected for immunohistochemical analysis using an anti–BrdU antibody at E18.5. The arrows indicate BrdU and GFP double-positive cells. Insets show high-magnification views. Bars: (main) 25 µm; (insets) 10 µm for. The bar graph exhibits the percentage of GFP and BrdU double-positive cells relative to the total number of GFP-positive cells in the CP (n = 3; mean ± SEM; **, P < 0.01; t test, two sided). (D–F) The protein levels of PAX6, SOX2, and TUJ1 were determined by Western blot analysis in mouse NPCs after the infection of control or HIRA-shRNA lentivirus. β-actin was used as a control. The bar graphs show the relative band intensity of PAX6 (E) and TUJ1 (F) in mouse NPCs (n = 3; mean ± SEM; **, P < 0.01; t test, two sided).