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. 2016 Nov 22;173(1):582–599. doi: 10.1104/pp.16.01602

Figure 2.

Figure 2.

Phospho-mutations at S532 and T552 alter MAP65-1 properties in vivo. A, Colocalization between Aurora1-GFP and MAP65-1-RFP throughout mitosis. The panels are taken from three different time-lapse movies. The fluorescence intensity plots (shown below the merged images) demonstrate colocalization of Aurora1-GFP and MAP65-1-RFP e.g. at the nuclear envelope during the PPB stage and during cytokinesis as well as a negative correlation between Aurora1 localization and MAP65-1 MT bundling from prophase to cytokinesis. Scale bar = 10 μm. B, Representative images (n > 10) of Arabidopsis MAP65-1-GFP, MAP65-1(AA)-GFP, and MAP65-1(DD)-GFP expressed in tobacco BY-2 cells during consecutive stages of cell division. Arrows indicate excessive microtubule bundling observed in prophase, metaphase, and late cytokinesis in cells expressing MAP65-1(AA)-GFP. Images of cells in interphase and preprophase are projections, while the other phases are represented by single images. Scale bar = 10 μm. C, Box plot representation of full cell division (top), metaphase (middle), and cytokinesis (bottom) duration in BY-2 cells expressing MAP65-1-GFP (n = 13), MAP65-1(AA)-GFP (n = 14), and MAP65-1(DD)-GFP (n = 9). Overexpression of MAP65-1(AA), but also MAP65-1(DD) prolongs cell division duration compared to the wild-type MAP65-1 (t test; triple asterisk: P < 0.0001; double asterisk: P < 0.001; ns (not significant): P > 0.01). Center lines show the medians; box limits indicate the 25th and 75th percentiles as determined by R software; whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles. D, Fluorescence intensity plots (taken along the white lines in B) across the phragmoplasts in early or late cytokinesis in BY-2 cells. MAP65-1(AA)-expressing cells show delayed depolymerization of central phragmoplast microtubules in late cytokinesis.