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. 2019 Mar 13;39(11):1994–2010. doi: 10.1523/JNEUROSCI.1849-18.2018

Figure 7.

Figure 7.

Cenpj regulates cortical development via Kif2a. A, ARPE19 cells were Immunostained with anti-γ-tubulin (gray) and anti-Cenpj (red) antibodies. Scale bars, 2 μm. B, ARPE19 cells were transfected with GFP-Kif2a and immunostained with anti-acetylated tubulin (gray) and anti-Cenpj (red) antibodies. Arrow indicates the cilium. Scale bar, 10 μm. C, Overexpressed Kif2a was efficiently knocked down by Kif2a shRNAs. GAPDH serving as a loading control. D, Quantification of Kif2a knock-down efficiency by shRNAs. Histograms show the mean ± SD; ****pshCtrl vs shKif2a-1 < 0.0001, ****pshCtrl vs shKif2a-2 < 0.0001 as determined by a t test; n = 3. E, Analysis of the radial migration of cortices 3 d after IUE at E13.5 with control shRNA, shKif2a-1, and shKif2a-2. Scale bar, 30 μm. F, Quantification of the neurogenesis after silencing Kif2a by measuring the percentages of GFP+ cells that have reached the different zones of the cortex 3 d after electroporation. Histograms show the mean ± SD; shCtrl vs shKif2a-2 (****pCP < 0.0001, **pIZ = 0.0025, ****pVZ/SVZ < 0.0001 as determined by t test; n = 3; 9 brain slices per experiment). G, Kif2a are specifically knocked down by shKif2a-2 and rescued by overexpression of Kif2a-td plasmid. GAPDH served as a loading control. H, Quantification of Kif2a protein expression index. Histograms show the mean ± SD; ****pshCtrl+RFP vs shKif2a-2+RFP < 0.0001 as determined by a t test; n = 3. I, Analysis of the radial migration of cortices 3 d after IUE at E13.5. Scale bar, 30 μm.