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. 2016 Jan 22;7:10481. doi: 10.1038/ncomms10481

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(a) qPCR analysis of the expression of genes related to the Shh signalling pathway in the cortex of Rcor2^fl/fl and Rcor2^cko brains during development. Significant upregulation of these genes on Rcor2 depletion is noteworthy. Transcripts were normalized to Rcor2^fl/fl group. Data are shown as mean±s.d., t-test, *P<0.05, **P<0.01 and ***P<0.001, n=3. (b) Confocal images of Shh and Ptch1 expressions in Rcor2^fl/fl and Rcor2^cko cortex. Enhanced Shh and Ptch1 signals are observed in Rcor2^cko compared with Rcor2^fl/fl neocortex at E13.5 and E15.5. Insets show high-magnification images of the outlined regions. Scale bars, 50 μm. (c) Dlx2 expression in Rcor2^fl/fl and Rcor2^cko cortex detected by immunofluorescence analysis at E15.5. Dlx2⁺ cells were observed in the neocortex on Rcor2 depletion. Scale bars, 50 μm. (d) Confocal images of Shh, Ptch1 and Dlx2 expressions in in-vitro-cultured Rcor2^cko NPCs. Scale bar, 20 μm. (e) Western blot analysis of expression levels of Dlx2, Shh and Ptch1 in Rcor2^fl/fl and Rcor2^cko cortex at E15.5. β-Actin is used as an endogenous control.