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Fig. 7 The phenotypic effect of destabilising Her6 protein is negligible in 24 hpf embryos but becomes more evident by 48 hpf. (A-C) qPCR quantification of mRNA expression level observed in elavl3 (A), ngn1 (B) and ascl1 (C) between HV (green) and HVP (pink) in dissected telencephalon of 24 and 48 hpf embryos. Boxes indicate median and interquartile range. Whisker endpoints indicate 25th percentile−1.5× interquartile range and 75th percentile+1.5× interquartile range. ns: P>0.05. (D,E) Representative examples of elavl3 (D) and ngn1 (E) mRNA visualised using HCR in HV and HVP forebrain at 24 and 48 hpf. Frontal view. Red arrows indicate telencephalic region. Yellow dashed line indicates embryonic outline. (F,G) Double HCR was performed on each embryo for elavl3 or ngn1 with foxg1. Relative volume was calculated as a ratio of the respective gene with foxg1. (F) Relative volume elavl3/foxg1 compared between HV and HVP embryos at different stages. (G) Relative volume of ngn1/foxg1 compared between HV and HVP embryos at different stages. (H-K) Quantification of mRNA by qPCR showing the expression level of pallial glutamatergic markers, vglut2a and tbr1 (H,I), and sub-pallial GABAergic markers, gad1 and gad2 (J,K). Boxes indicate median and interquartile range. Whisker endpoints indicate 25th percentile−1.5× interquartile range and 75th percentile+1.5× interquartile range. ns: P>0.05. (L,M) Representative examples of vglut2a (M) and gad1 (N) mRNA visualised using HCR in HV and HVP forebrain at 24 and 48 hpf. Frontal view in L and M, 48 hpf. Lateral view in M, 24 hpf. Red arrows indicate telencephalic region. Yellow dashed line indicates embryonic area. (N,O) Double HCR was performed on each embryo for vglut2a or gad1 with foxg1. Relative volume was calculated as a ratio of the respective gene with foxg1. (N) Relative volume of vglut2a/foxg1 compared between HV and HVP embryos at different stages. (O) Relative volume of gad1/foxg1 compared between HV and HVP embryos at different stages. *P≤0.05; **P≤0.01; ***P≤0.001; ****P≤0.0001 (statistical test details found in Table S6). Scale bars: 50 µm.