Additional file 1 of A heat-sensitive Osh protein controls PI4P polarity

Figure S1. PI4P distribution is regulated by growth conditions. (a) Cells expressing the PI4P reporter GFP-P4C (green) and a PM marker mCherry-2xPHPLCδ (magenta) were grown at 26 °C (left panels) and subjected to a heat shock for 10 min at 42 °C (right panels). Mother cells are indicated (m) and arrows point to GFP-P4C localization at the PM of mother cells at 42 °C. Scale bars, 5 μm. (b) Representative images of a time course of cells expressing the PI4P reporter GFP-P4C subjected to a heat shock at 42 °C. Cells were grown at 26 °C, immobilized on a 2% agarose pad mounted on a microscope slide. Arrowheads point to GFP-P4C localization at the PM of mother cells. Cells were imaged over time at 42 °C using a BIOPTECHS Objective Heater System. Scale bar, 5 μm. (c) Graph displays the mean Fd/Fm ratios of GFP-P4C fluorescence at 26 °C (t=0) and during heat shock at 42 °C for different time points (2 min intervals, see B). Error bars represent standard deviation. In total, 10 cells from two independent experiments were analyzed. (d) Graph shows the mean GFP-P4C fluorescence intensity at the mother cell PM (Fm) at 26 °C (t=0) and during heat shock at 42 °C at different time points (2 min intervals, see b and c). In total, 10 cells from two independent experiments were analyzed.