Fig. 3.

AtPH1 localization depends on binding to PI3P. (A) Confocal images of the root elongation zone of nns1 plants expressing AtPH1-GFP or AtPH1R46H-GFP under the control of the 2x35S promoter. The boxed area in the left panel is enlarged at right. Cell walls stained by PI are in magenta. The white arrow indicates the vacuolar membrane detaching from the cell periphery. (B) nns1 plants expressing AtPH1-GFP were crossed with plants expressing the PI3P marker 2xFYVE-mCherry. Colocalization of both fluorescent markers was analyzed by confocal microscopy on roots of F1 plants. On the merged picture the overlap of GFP (green) and mCherry (magenta) channels appears in white. Cell contours are represented by dashed lines. Pearson (r_p) and Spearman (r_s) correlation coefficients as well as M1 (GFP) and M2 (mCherry) Manders overlap coefficients above threshold were calculated. (C) The roots of nns1 plants expressing AtPH1-GFP were treated by 30 µM wortmannin or 0.1% DMSO for 3 h. (Scale bars: 10 µm.)