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. 2017 Jul 6;7:4787. doi: 10.1038/s41598-017-05305-0

Figure 1.

Figure 1

Effect of Znf179 on neuronal differentiation. (A,B) Knockdown of Znf179 expression diminished neuronal differentiation and dendritic arborization in primary cerebellar granule cells. Primary culture of cerebellar granule cells from cerebella of wild-type or Znf-179-knockout mice were stained with anti-MAP2 (red) and DAPI (white). The Quantification of the percentage of MAP2(+) cells is shown in the histogram. **p = 0.0018. Scale bar, 50 μm. (n > 300, groups were compared using t-test, two-tailed p value). (C,D) N2a cells with and without stable expression of GFP-Znf179 proteins were induced to differentiate in MEM containing 2% serum for 24 h. The overexpression of GFP-Znf179 induced N2a cell differentiation in both 10% and 2% serum-containing medium, while those in 2% serum-containing medium exhibited longer neurite lengths (the lower right panel in 1 C). Significant neurite branching was observed in differentiated N2a cells. Differentiated and total N2a cells were determined in at least five fields of views, and at least 600 cells were counted in total. Quantification of the percentage of differentiated N2a cells is shown in the histogram. ***p < 0.001. Scale bar, 50 μm. (n > 600, groups were compared by a one-way ANOVA, two-tailed p value). (E) Total cell lysates of N2a cells were analyzed by Western blotting using anti-MAP2, anti-Znf179, anti-Tuj-1, anti-GFAP, anti-α-tubulin and anti-GAPDH antibodies. Uncropped images are in Supplementary Figure S2.