Extended Data Fig. 4: Augmin and TPX2 spindle localization is comparable between the differentiation states.
From: Cell state-specific cytoplasmic density controls spindle architecture and scaling
a. Cellular levels of augmin (subunit HAUS6) after 48 h of differentiation relative to ESCs, probed by western blotting and normalized to tubulin (N = 3 biological replicates). Bars show the mean, errors show the standard deviation, circles show replicates. Welch’s t-test (two-sided), P = 0.57. b. Confocal micrographs (maximum projected, N = 2 experiments) of fixed ESCs or DIF at metaphase. Top: Immunostained HAUS6 signal, bottom: tubulin::GFP (grey) and chromatin counterstained by Hoechst (blue). Dotted lines indicate the cell boundaries. Scale bar: 5 µm. c. Fluorescent HAUS6 signal on spindle (normalized by total cell HAUS6) as a function of spindle volume. Each data point represents a single cell (ESCs n = 159 cells and DIF n = 158 cells pooled from 2 independent experiments), large circles denote medians in each cell volume bin, error bars show interquartile ranges. d. As in a) but showing TPX2 levels (N = 3 biological replicates). P = 0.92. e. As in b) but showing TPX2 immunofluorescence (N = 3 experiments). f. As in c) but showing TPX2 immunofluorescence (ESCs n = 105 cells and DIF n = 141 cells pooled from 3 independent experiments). n.s.: not significant, P > 0.05.
