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. 2016 Feb 1;143(3):398–410. doi: 10.1242/dev.130849

Fig. 6.

Fig. 6.

WNT-induced human NC cells arise independently of PAX6+ neuroepithelial cells. (A) Immunostaining of OCT4 (violet), SOX10 (green) and PAX6 (red) in day 1, day 3 and day 5 cultures treated with (+) or without (−) CHIR. Framed regions, from day 3 (Aa, Ab) and day 5 (Ac) cultures are shown as magnified images; arrows highlight the few PAX6+ cells in day 3 +CHIR cultures. (B) Quantification of SOX10+ and PAX6+ nuclei in day 1, 3 and 5 cultures treated with CHIR. (C) Western blot of PAX6 protein in control and PAX6 shRNA-treated cells, with ladder sizes indicated. (D,E) Quantitative gene expression analysis showing effective knockdown of PAX6 at day 3 (D), but no reduction of NC markers at day 5 in PAX6-specific shRNA-treated cells (E). *P≤0.05, ***P≤0.001, ****P≤0.0001. Error bars represent s.d.