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Fig. 5. Fezf2 represses the activity of lhx2 and lhx9 in the forebrain. (A) ChIP-qPCR analysis of Fezf2 binding to the lhx2 promoter. Region 1 showed very high enrichment (n=3 replicates). (B,C) qPCR analysis of lhx2 and lhx9 expression in p3hGR-VP16-Fezf2-injected animal cap explants aged to stage 12 and treated with CHX alone or CHX+DEX (n=3 replicates). (D,E) qPCR analysis of lhx2 and lhx9 expression in neuralised animal cap explants aged to stage 20 (n=3 replicates). (F,G) In situ hybridisation analysis shows that mild knockdown of fezf2 leads to expansion of the lhx2 (F) and lhx9 (G) expression area. Arrowhead indicates the epithalamus; bracket indicates the ventral diencephalon. (H,I) In situ hybridisation analysis of arx (H) or ngn1 (I) in stage 28 morphants (lateral views). Arrowheads indicate arx or ngn1 expression. (J) qPCR analysis shows that lhx2 and lhx9 antagonise expression of the fezf2-induced Wnt-responsive gene xnr3 in stage 14 animal cap explants (n=3 replicates). (K) qPCR analysis shows lhx2 and lhx9 antagonise fezf2-induced ngn1 expression in stage 20 animal cap explants (n=3 replicates). In all qPCR analyses, rpl8 was used as internal control. Error bars represent s.e.m. *P<0.05, ***P<0.001; ns, not significant. |
