Extended Data Fig. 1: Marker expression and tissue architecture confirm successful differentiation of ESCs into neural rosettes.
From: Cell state-specific cytoplasmic density controls spindle architecture and scaling
a. Top: Confocal data showing immunostaining of PAX6109 in fixed ESCs (“Day 0”) and during 5 days of neural differentiation. Bottom: Nuclei stained by Hoechst. Scale bar: 50 µm. b. Immunofluorescent detection of nestin110 in ESCs-derived rosette cells after 5 days of differentiation. Inlets show nuclei stained by Hoechst. Scale bars: 20 µm. c. As in (b) but detecting MASH1 (also known as ASCL1), a proneural transcription factor111. d. As in (b) but detecting ZO-1, marking tight junctions in the rosette centre112. e. As in (b) but detecting PAX6, uniformly expressed in the rosette bulk. f. As in (b) but detecting TBR2113, sporadically expressed by cells in the rosette periphery. g. As in (b) but detecting AKNA, a centrosomal protein in neural stem and progenitor cells. AKNA+ centrosomes strictly localize to the “apical” pole during interphase114. Lookup table of this panel is inverted in comparison with other panels for better visibility. All stainings (a-g) were performed for N = 2 independent experiments.
