Localization of PtdIns(4)P, PtdIns(4,5)P2 or PtdIns(3,4,5)P3 in PC12 cells carrying scrambled shRNA or <i>VPS13A</i> knockdown shRNA.

<p>PC12 cells were transiently co-transfected with GFP tagged PH domains and vectors expressing either a control, scrambled shRNA or <i>VPS13A</i> shRNA3. Red fluorescence indicates the presence of the shRNA vector. For each sensor, the percentage of knockdown cells in which the sensor was localized at the plasma membrane and the number (n) of cells scored are shown. The data are pooled from three experiments for the PtdIns(4)P and PtdIns(4,5)P₂ sensor and two experiments for the PtdIns(3,4,5)P₃ sensor. In <i>VPS13A</i> knockdown cells, three patterns of localization for the PtdIns(4)P sensor are seen: class I are similar to the control cells (indicated by “I”); class IIa are lacking in plasma membrane fluorescence but have clear Golgi and nuclear fluorescence (IIa); and class IIb show diffuse fluorescence throughout the cell (IIb). Yellow arrows indicate the plasma membrane. White arrowheads indicate Golgi elements. N = nucleus. Scale bars = 5μm.