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. 2022 Sep 6;20(9):e3001563. doi: 10.1371/journal.pbio.3001563

Fig 7. Foxg1 required cell autonomously for eGC production.

Fig 7

(A) A hypothesis: Pax6 suppresses specifically Foxg1-permitted Shh-induced generation of eGCs without interfering with other effects of Shh pathway activation in these cells. (B) The experimental procedure for (C-E): TAM was administered at E9.5 to generate Pax6 cKOs in which neither, one, or both Foxg1 allele(s) were also deleted (for alleles, see S16A Fig); brains were analysed at E13.5, E14.5, or E16.5. (C) Immunoreactivity for Gsx2 and GFP and in situ hybridizations for Gad1 and Dlx1 in E14.5 cortex from Pax6 cKO embryos in which neither, one, or both Foxg1 allele(s) were deleted by TAM at E9.5. Scale bar: 0.1 mm. (D) Densities of Gsx2+ cells in the lateral cortex of E14.5 embryos with the 3 genotypes in (C) (averages ± SEM; n = 3 embryos of each genotype, from 3 independent litters) (Sheet C in S7 Data). (E) High magnification images from (C): At least the majority of residual Gad1+ (green) cells in Pax6 Foxg1 double KOs (arrows) were GFP-negative (i.e., not white) of subcortical origin (non-Emx1-lineage). Scale bar: 0.01 mm. (F) The experimental procedure for (G): Pax6 Foxg1 dcKO cortex made using Emx1-Cre, avoiding the need for TAM; a construct expressing Foxg1 and mCherry was electroporated into the cortex on E14.5; coronal sections were analysed on E16.5. (G) Results of experiment in (F): expression and coexpression of Foxg1, Gsx2, Olig2, mCherry, and GFP protein and Dlx1 mRNA in coronal sections. Scale bars: 0.1 mm. dcKO, double conditional KO; eGC, ectopic GABAergic cell; GFP, green fluorescent protein; Pax6 cKO, Pax6 conditional knockout; TAM, tamoxifen.