Figure 3.

Lineage-Reprogrammed Cortical and Cerebellar iNs Adopt Different Neurotransmitter Phenotypes

(A–J) Examples of CerebAstro-derived RFP⁺ iNs expressing GABA 20 dpn with either ASCL1 (A) or NEUROG2 (D). Single confocal z stacks are shown in higher magnification to confirm the colocalization of GABA and RFP in iNs (B, C, and E–J). Observe the similar pattern of GABA expression between co-cultured hippocampal neurons (RFP⁻, yellow arrowhead in D) and iNs (RFP⁺, white arrowheads in D).

(K–R) Example of CerebAstro-derived RFP⁺ iNs expressing GLUT 20 dpn with either ASCL1 (K–O) or NEUROG2 (P–R). Single confocal z stacks are shown in higher magnification to confirm the colocalization of glutamate and RFP in iNs (B, C, and E–J). Observe the presence of GLUT⁻ (K–M, yellow arrowhead) and GLUT⁺ (K, N, and O, white arrowhead) RFP⁺ iNs.

(S) Quantification of GABA⁺ and GLUT⁺ cells among RFP⁺ iNs derived from CerebAstro nucleofected with ASCL1 (orange) or NEUROG2 (yellow) and CtxAstro nucleofected with ASCL1 (dark green) or NEUROG2 (light green). ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001.

(T–Z′) Expression of GFP in CerebAstro and CtxAstro derived from GAD67-GFP mice 12 dpn with ASCL1. Example of RFP⁺/GFP⁺/BIII-TUB⁺ iN derived from CerebAstro (T, dashed box magnified and showing single channels in U–W) or CtxAstro (X, dashed box magnified and showing single channels in Y–Z′). GLUT, GLUTAMATE; BIII-TUB, BIII-TUBULIN. Statistical test: two-way ANOVA followed by Bonferroni’s post hoc test (mean ± SEM).

Data are derived from 3 independent experiments. Scale bars, 25 μm. See also Figure S5.