Fig. 3: Single-cell sequencing of H9 and 4X progenitors at DIV16 using a hindbrain differentiation protocol. | Nature Communications

Fig. 3: Single-cell sequencing of H9 and 4X progenitors at DIV16 using a hindbrain differentiation protocol.

From: Enhanced production of mesencephalic dopaminergic neurons from lineage-restricted human undifferentiated stem cells

Fig. 3

a Diagram of hindbrain differentiation protocol. b Graph of the cluster composition for H9 and 4X at DIV16. c Graph showing abundance of H9 and 4X cells in each cluster at DIV16. d UMAP of H9 and 4X cells at DIV16 (H9 cell spheres: n = 10; 4X cell spheres: n = 10; total of 4682 cells). e Heatmap of selected genes expressed in each cluster. f Feature plot of the contribution of each cell line to each cluster and feature plot of gene expression levels of OTX2, EN1, LMX1A, FOXA2, FGF8, HOXA/B family members and STMN2. g Percentage of H9 and 4X cells expressing OTX2 and EN1. h Representative immunofluorescence analysis of OTX2/EN1, OTX2/FOXA2 and FOXA2/LMX1A expression in H9 and 4X cells treated with 1 µM GSK3i on DIV16, n = 3 independent experiments. DAPI was used as a nuclear stain. Scale bars, 50 µm. Source data are provided as a Source Data file.

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