Figure 2.

GATA6^4ins/+ and GATA6^Δ4/Δ4 Mutant hESC Lines Display Impaired DE Formation

(A) Expression of pluripotency (OCT4, SOX2), primitive streak (BRACHYURY), mesendoderm (EOMES), and definitive endoderm (CXCR4, SOX17, GATA4) markers, as well as GATA6 itself, in H9^∗ and H9-derived GATA6^4ins/+ and GATA6^Δ4/Δ4 mutant cells over 3 days of differentiation (Figure S1A).

(B) Differentiation efficiency measured by FACS analysis of CXCR4 and SOX17 at day 3 DE in H9^∗ and H9-derived GATA6^4ins/+ and GATA6^Δ4/Δ4 mutant cells.

(C) Immunofluorescence analyses for the key DE markers GATA6 with SOX17 in H9^∗ and H9-derived GATA6^4ins/+ and GATA6^Δ4/Δ4 mutant cells. DAPI, 4′,6-diamidino-2-phenylindole. Scale bars, 100 μm.

(D) Expression of pluripotency (OCT4) and definitive endoderm (SOX17, CXCR4) markers in H9^∗ and H9-derived GATA6^4ins/+ and GATA6^Δ4/Δ4 mutant cells on days 3 and 6 of differentiation with the STEMdiff pancreatic progenitor kit.

(E) Differentiation efficiency measured by FACS analysis of CXCR4 and SOX17 at day 3 DE in H9^∗ and H9-derived GATA6^4ins/+ and GATA6^Δ4/Δ4 mutant cells differentiated using the STEMdiff pancreatic progenitor kit.

(A and D) Error bars represent the SE of three independent experiments. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, ^∗∗∗∗p < 0.0001.

(B and E) Undifferentiated hESCs stained with the respective primary and secondary antibodies and secondary antibody only (IgG) were both used as controls. Gates were set according to an hESC control.