Figure 1.

Defining diversity of interneurons generated via TF-induced programming of human PSCs

(A) Schematic of the ASCL1/DLX2 expression constructs and procedure used to generate interneurons (iINs) with the AD method.

(B) Immunostaining of iINs after 2 weeks for HuNu, TUJ1, and GABA.

(C) qPCR for pluripotency- and interneuron-specific transcripts in iINs. Error bars are representative of three technical repeats.

(D) Representative patch clamp analysis of two iINs 2 weeks after induction, measuring action potential frequency. Membrane voltage traces (top) show the effects of depolarizing (black) and hyperpolarizing (red) current injections. The traces of the injected currents (duration 200 ms) are shown in bottom panels. Note the different modes of firing in the two cells: (left) repetitive firing and (right) only one action potential fired upon depolarization and a rebound action potential following the hyperpolarizing pulse.

(E) Quality analysis of scRNA-seq data for the two biological replicates of iINs at 2 weeks of induction (after dox withdrawal), measuring the number (No) of transcripts and genes, and percentage of mitochondrial reads per cell across.

(F) UMAP clustering of single iINs for the two repeats in (E).

(G) Quality control showing even distribution of cells from both replicates among the eight clusters from (F).

(H) Normalized expression levels of indicated genes superimposed onto the UMAPs from each replicate. Colored according to expression levels by cell.

(I) As in (H), except for the interneuron subtype marker, SST.

(J) As in (I), except for cells that express both SST and CALB2 (left, in blue) or SST and NPY (right, in red).