Additional western blots and controls.

<p>(<b>A</b>) Phospho-Rps6 and phospho-Psk1 levels <i>in WT</i> heterothallic <i>(</i>h-<i>)</i> and homothallic <i>(</i>h90<i>)</i> cells starved for 6 h 15 and then treated with Torin1 (5 or 25 μm) or DMSO (0) for 30 min. (<b>B</b>) CFP-Atg8 cleavage in WT and <i>atg1Δ</i> mutants after 4 h and 8 h of nitrogen starvation in MSL. The cells were pre-grown in MSL + glutamate (top) or MSL + ammonium (bottom). (<b>C</b>) Time course of phospho-Psk1 in <i>h- sxa2Δ atg18aΔ</i> cells transferred to MSL + 0.75 mg/ml glutamate in the presence of P-factor (1 μg/ml) or MetOH at T = 0 min. CFP-Atg8 cleavage is shown in the bottom blot. (<b>D</b>) Time course of CFP-Atg8 cleavage in <i>h- sxa2Δ</i> cells, which are otherwise WT (top), <i>atg1Δ</i> (middle), or <i>atg5Δ</i> (bottom), transferred to MSL + 0.75 mg/ml (or 0.5 mg/ml in case of <i>atg1Δ</i>) glutamate in the presence of P-factor (1 μg/ml) or MetOH at T = 0 min. These are the same extracts as probed for phospho-Psk1 in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3002963#pbio.3002963.g006" target="_blank">Fig 6F</a>. Uncropped western blots available in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3002963#pbio.3002963.s012" target="_blank">S1 Raw Images</a>.</p> <p>(PDF)</p>