Extended Data Fig. 6: iDePP enables efficient depletion of PI(4,5)P2 in various tissues and organs.
From: Inducible depletion of PI(4,5)P2 by the synthetic iDePP system in Arabidopsis
a-c, Subcellular localization of mCIT-2xPHPLC and either MAP-mCH-dOCRL (a), or MAP-3xmCH; N = 12 (b), MAP-mCH-dOCRLdead; N = 12 (c), in cotyledon epidermal cells; N = 6. Fluorescence intensity is color-coded (green fire blue scale), cytoplasmic strands are indicated by the arrows. Scale bars, 10 µm. d-g, Induction of the iDePP system in the shoot apical meristem of Arabidopsis; N = 15 (Scale bar, 20 μm in (a-b), 10 μm in (c-d). Asterisks represent the position of the nucleus in the cell. (h) and (i) Subcellular localization of MAP-mCH-dOCRL (left panel) and mCIT-2xPHPLC (right panel) in a naked shoot meristem from an NPA-treated seedling, as viewed from the top; N = 4. In the absence of dex induction, mCIT-2xPHPLC was localized at the plasma membrane throughout the meristem, including both the central and peripheral zones (h, right panel). By contrast, upon induction of MAP-mCH-dOCRL expression, mCIT-2xPHPLC was no longer sharply localizing at the cell edge (i, right panel). Instead, mCIT-2xPHPLC accumulated in the cytosol, while still being excluded from the round central nuclei (i, right panel). Note that after dex induction, MAP-mCH-dOCRL was strongly expressed in the peripheral zone, and was only weekly expressed in the central zone (i, left panel). However, although its expression was weak in the central zone it appeared to be sufficient to impact mCIT-2xPHPLC localization. Scale bar, 20 μm. (j) and (k) Effect of PI(4,5)P2 depletion on organ initiation in NPA-treated meristems; N = 8. Altogether, these observations suggest that PI(4,5)P2 depletion seems to affect organ initiation in NPA-treated meristems. Scale bars, 700 µm.
