Extended Data Figure 9. The PHO1 transporter gene family as marker for passage cells.

a) Expression of PHO1 family members in the cortex/epidermis (left panel) and inside the stele (right panel) within the suberised zone of 5-day-old roots grown on ½ MS media. Individual nuclei showing expression of the respective NLS reporter were counted in similar 3D stacks taken in either a fully suberized area (red bars) or an area containing one passage cell (white bars). Open circles depict outliers. b) Expression of PHO1 in 5-day-old seedling in the late endodermis of Wildtype or a suberin-degradation line with virtually absent suberin (pCASP1::CDEF1). c) Representative images of each of the PHO1 members’ expression, highlighted by NLS-3Venus reporter, found to be expressed in roots of 5-day-old seedlings. Seedlings stained with Nile Red for suberin and Calcofluor White for cell walls. Scale bars in XZ projections represent 10 μm. White arrowheads highlight passage cell nuclei, yellow arrowheads point toward nuclei of cortex/epidermal cells. Asterisks emphasize passage cells. nd; not detected. Boxplot centers show median. ** depicts significant differences between groups (P<0,01) according to a two-sided T-test. For more information on data plots see the statistics and reproducibility section. In b) and c) the images represent 15 and 11 independent lines respectively, all stainings were repeated 3 times. n represents independent biological samples. Unless otherwise stated, scale bars represent 25 μm. For inserts scale bars represent 10 μm.