Fig. 6.
Colocalization of BIN2 with different OPS variants. (A–G) Transient expression of indicated mRFP (red fluorescence) or CITRINE (green fluorescence) fusion proteins in tobacco (N. benthamiana) leaf epidermal cells under the control of the 35S constitutive promoter (confocal microscopy). (A, Inset) Arrowhead points out nuclear localization. (H–M) Transient coexpression of mRFP-BIN2 with indicated CITRINE fusions of OPS variants (H′–M′) in tobacco leaf epidermal cells. Red fluorescent and green fluorescent channels are shown separately and in overlay (H′′-M′′). The number of cells that displayed the nuclear mRFP-BIN2 signal among all cotransformed cells is indicated in the lower right corner of H–M [compare with A; no combination behaved significantly differently from BIN2 + OPS, except BIN2 + OPSδ2−54 (P < 0.001), which, unlike all other combinations, was not significantly different from the BIN2 control (Fisher’s exact test)]. (H′′′–M′′′) Pearson’s r of colocalization (±SEM) (n ≥ 15 per combination). Expression levels (mean intensity value of transformed cells) of BIN2 or OPS variant fusion proteins were not significantly different within experiments (one-way ANOVA; point variants experiment: F = 0.21, P = 0.81 for OPS variants; F = 0.57, P = 0.56 for BIN2; deletion variants experiment: F = 2.357, P = 0.10 for OPS variants; F = 1.61, P = 0.21 for BIN2).