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. 2024 Feb 12;121(8):e2310502121. doi: 10.1073/pnas.2310502121

Fig. 5.

Fig. 5.

GATA2 and GATA3 are essential regulators for SynT development in humans. (A) Control, GATA2 KD, and GATA3 KD human TSCs were subjected to 2D SynT differentiation on collagen-coated adherent cell culture dishes. Image panels show impaired ST(2D) colony formation in GATA2 KD and GATA3 KD cells. (Scale bars, 200 μm.). (B) Immunofluorescence analysis of the ST(2D) colonies (Left) marked by the loss of E-Cadherin (CDH1, green) expression but induction of HCGβ expression (red). GATA3 KD ST(2D) show altered cellular morphology, maintenance of E-Cadherin (CDH1, green) expression, and impaired induction of HCGβ expression (Right). GATA2 KD ST(2D) colonies (Middle) were much smaller; there was impaired cell fusion as E-Cadherin was maintained compared to the control but lacked HCGβ expression. (C) Immunofluorescence analysis of the ST(3D) colonies revealed loss of E-Cadherin (CDH1, red) expression and induction of HCGβ expression (green). Both GATA2KD and GATA3 KD ST(3D) show altered cellular morphology, maintenance of CDH1 expression, and impaired induction of HCGβ expression. (D) Quantitative RT-PCR analyses (mean ± SE; n = 3, P ≤ 0.001) reveal impaired induction of SynT markers like CGB, ERVW-1, PSG4, CGA, and SDC-1 in GATA2 KD and GATA3 KD human TSCs, undergoing 3D SynT differentiation.