Fig. 4: ANXA1 is required for correct mitotic progression and spindle dynamics. | Nature Communications

Fig. 4: ANXA1 is required for correct mitotic progression and spindle dynamics.

From: Annexin A1 is a polarity cue that directs mitotic spindle orientation during mammalian epithelial morphogenesis

Fig. 4

a Time-lapse images of representative MCF-10A cells treated with si-Control or si-ANXA1#1. Microtubules are labelled with SiR-tubulin (green) and DNA with Hoechst 33342 (magenta), 2 h and 30 min before acquisition, respectively. Arrowheads indicate the phenotypes listed in (e). Scale bars, 5 µm. b Percentage of cells with abnormal mitosis (si-Control: n = 33; si-ANXA1#1: n = 44). Two-sided t-test, ****P < 0.0001. c Percentage of cells that completed mitosis (si-Control: n = 33; si-ANXA1#1: n = 44). Two-sided t-test, **P = 0.0095. d Time from nuclear envelope breakdown (NEBD) to anaphase onset in siRNA-transfected cells (si-Control: n = 33; si-ANXA1#1: n = 25). Two-sided t-test, **P = 0.006. e Percentage of chromosomes segregation and spindle assembly defects in siRNA-transfected cells (si-Control: n = 33; si-ANXA1#1: n = 44). Two-sided t test, normal segregation: ****P = 0.000033; chromosome bridges: **P = 0.0055; abnormal bipolar spindle: **P = 0.0043; multipolar spindle: *P = 0.032. f Dynamics of z orientation (αz) during metaphase in siRNA-transfected cells (si-Control: n = 33; si-ANXA1#1: n = 39). Black lines represent the average spindle angles. g Oscillation index in siRNA-transfected cells, in the z-axis (si-Control: n = 33; si-ANXA1#1: n = 41). Two-sided t-test, *P = 0.0125. h Percentage of cells with spindle oscillations in the z axis (si-Control: n = 33; si-ANXA1#1: n = 44). Two-sided t-test, ****P < 0.0001. i Dynamics of xy orientation (αxy) during metaphase in siRNA-transfected cells (si-Control: n = 33; si-ANXA1#1: n = 39). Black lines represent the average spindle angles. j Oscillation index in siRNA-transfected cells, in the xy axis (si-Control: n = 33; si-ANXA1#1: n = 41). Two-sided t-test, *P = 0.0153. k Percentage of cells with spindle oscillations in the xy axis (si-Control: n = 33; si-ANXA1#1: n = 44). Two-sided t test, *P = 0.0105. l Origin-aligned metaphase plate tracks in the xy plane of si-Control- (left) and si-ANXA1#1-transfected (right) cells (si-Control: n = 33; si-ANXA1#1: n = 43). m Displacement length from the starting point to the position at the end of metaphase in siRNA-transfected cells (si-Control: n = 31; si-ANXA1#1: n = 30). Two-sided t-test, *P = 0.037. n Velocity of metaphase plate movements in si-RNA-transfected, calculated by dividing the track length by metaphase duration (si-Control: n = 31; si-ANXA1#1: n = 44). Two-sided t-test, P = 0.384. All data are presented as mean ± s.e.m. from 3 independent experiments. Source data are provided as a Source Data file.

Back to article page