Figure 4.

Validation of marker expression in hypothalamus organoids

(A) qRT-PCR of hPSCs and D86 organoids for several markers identified by scRNA-seq analysis (n = 3). ACTB was used as a housekeeping gene for normalization. Error bars represent SD. qRT-PCR, quantitative real-time PCR.

(B) Western blot of hPSCs and D85 organoids for TH, NKX2-1, and OTP. GAPDH was used as loading control.

(C) IHC staining of D80 organoids for the astrocyte markers, GFAP and SOX9, as well as the glutamatergic neuron marker, VGlut2 (encoded by the SLC17A6 gene). Hematoxylin was used as a nuclear marker. Scale bars: 50 μm.

See also Figure S9.