DIDO1 and DIDO3 Expression in Early ESC Differentiation
(A) Left: immunostaining of WT day-4 embryoid bodies (d4EBs) for OCT4 (green) and GATA4 (red); arrowheads mark outer primitive endoderm (PE) cell layer. Right: lack of PE formation in Dido3ΔCT d4EBs.
(B) Left: localization of DIDO isoforms using anti-DIDO MAB-1C6 (green; mainly in nucleus) and DIDO3-specific PAB-DIDO3 (red). Right: localization of DIDO isoforms using MAB-1C6 (green) and DIDO1-specific PAB-DIDO1 (red; preferentially at the apical membrane of outer cell layer).
(C) Left: developing PE and apical localization of DIDO1 (arrowheads) in a time course of WT d1–d4EBs labeled for OCT4 (green) and DIDO1 (red) expression. Right: lack of PE formation in a time course of Dido3ΔCT d1–d4EBs labeled for OCT4 (green) and DIDO1 (red).
All panels are confocal images with nuclear counterstaining (DAPI, blue). Scale bars, 50 μm.