Figure 1. GATA6 and GATA4 expression during early pancreatic progenitor differentiation in hPSCs.

(A) Schematic of directed endoderm and PDX1+ early pancreatic progenitor differentiation from HUES8 hPSCs using the 1st-generation protocol. The cell types and key markers at each stage are shown. Chemicals and durations for each differentiation stage are indicated. PS: undifferentiated hPSC stage; DE: definitive endoderm stage; PP1: early pancreatic progenitor stage (cells expressing PDX1 but not NKX6.1); Activin A, a TGF-β superfamily ligand; BIO: BIO-acetoxime, a GSK3 inhibitor that activates WNT signaling; FGF10: fibroblast growth factor 10; SANT-1, a Hedgehog pathway antagonist; RA: retinoic acid; LDN: LDN-193189, a BMP type 1 receptor (ALK2/3) inhibitor. The day when differentiation is initiated is designated as day 0 (d0).

(B) The mRNA expression patterns of GATA6 and GATA4 during hPSC differentiation to early pancreatic progenitor. The mRNA levels were measured by RT-qPCR (n=4) and normalized to internal control ACTB. Student’s t-test with two-tailed distribution and two-sample equal variance was used for statistics analysis. P values <0.05, 0.01, and 0.001 were indicated with 1, 2, and 3 asterisks, respectively, in all figures.

(C) Representative immunostaining of GATA6 or GATA4 with other stage-specific markers including OCT4, SOX17, and PDX1. Scale bar, 100 μm for all images throughout.

(D) Representative FACS dot plots of cells stained for CXCR4, SOX17, GATA6 and GATA4 factors at the DE stage. The percentage of each cell population is indicated in the corresponding quadrant for all FACS plots.

(E) Representative FACS dot plots for co-expression of GATA6 or GATA4 with PDX1 at the PP1 stage.

All data in this figure were generated from HUES8 line using 1st-generation differentiation protocol. See also Method Details.