Fig. 5: Temporal RNA-seq of RGC-iNs. | npj Regenerative Medicine

Fig. 5: Temporal RNA-seq of RGC-iNs.

From: Human retinal ganglion cell neurons generated by synchronous BMP inhibition and transcription factor mediated reprogramming

Fig. 5

a Timeline for longitudinal studies. b RNA-seq PCA plot showing clustering among PSCs, days 6, 14, 21, and 28 (D6, -14, -21, -28) RGC-iN cultures and days 6, 14, and 21 (D6, -14, -21) CTLs. c Venn Diagram showing overlapping genes at each time point (normalized count > 10). d POU4F1-mNeonGreen/POU4F2-tdTomato dual reporters in RGC-iNs at day 6. e Cryosectioned day 45 organoids exhibiting POU4F2-tdTomato + /POU4F1-mNeonGreen+ reporter expression. f Heatmap indicating the expression of selected markers for housekeeping (HKG), pluripotency, NAIP2 cassette, photoreceptor (PRs), glial, RGC, inhibitory, excitatory, and pan-neuronal genes. g Line graph showing the RGC transcription factors (TFs) NEUROG2, ATOH7, ISL1, POU4F1, and POU4F2 expression at each time point. h Line graph for selected neurotrophin receptors and immunostaining of day 6 RGC-iNs with pCREB after 30 min BDNF treatment. i Heatmap highlighting ion channel genes expressed at each time point. j UMAPs of RGC-iNs colored by time point (top) and Louvain clusters (bottom). k UMAPs of RGC-iNs colored by pan-neuronal marker gene expression (top) and RGC-specific marker genes (bottom). l Heatmap showing normalized expression of various RGC-subtype marker genes in each cluster of RGC-iNs across day 7, 14, and 21 time points. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns P > 0.05, n = 3 for PSCs, n = 4 for all other samples. Error bars g, h = SEM. Scale d = 50 µm, e = 200 µm, e, inset = 50 µm, h = 85 µm.

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