a, Transient expression in tobacco leaf epidermis of BIN2–GFP, POLAR–mCherry, mCherry–BASL, and the kinase-dead version of BIN2 (BIN2(K69R)–GFP). b, Transient co-expression of BIN2–GFP or BIN2(K69R)–GFP with POLAR–mCherry excluding BIN2 from the nucleus. c, Transient co-expression of BIN2–GFP with mCherry–BASL excluding BASL from the nucleus, whereas the transient co-expression of BIN2(K69R)–GFP with mCherry–BASL did not. The nucleus is indicated with arrowheads (a–c). d, Quantification of the nuclear exclusion of BIN2 and BASL. n, number of cells from three biologically independent leaves. Images were quantified with the nuclear (N)-TagBFP2 as a marker for the nucleus area and by means of a script to measure the average intensity in the nucleus compared to the remainder of the cell (see Methods). For a–c, experiments were repeated independently three times. Box plots show the first and third quartiles, split by the median (line) and mean (cross). Statistical analysis was performed with a one-way ANOVA and Tukey’s post hoc test compared to BIN2–GFP or BIN2(K69R) or mCherry–BASL single infiltration, respectively. Scale bars, 20 µm.