<p><i>A</i>, the level of Sec14 and Sec14<sup>G266D</sup> in <i>sec14<sup>ts</sup> rpn4</i>Δ cells. <i>B</i>, the level of Sec14 and Sec14<sup>G266D</sup> in <i>sec14<sup>ts</sup></i> cells treated with MG132. Strains were transformed with plasmids expressing Sec14 or Sec14<sup>G266D</sup> containing an N-terminal T7 epitope tag and were grown to mid-logarithmic phase at 25°C, with a subset shifted to 37°C for 2 hours (A). For MG132 treatment cells were grown as before and shifted to 37°C in the presence of 100 μM MG132 for 2 hours. Cells were disrupted by three passes through a French press and unbroken cells removed by centrifugation. Protein extract was separated by SDS-PAGE, transferred to PVDF membrane, and western blots versus the T7 epitope were performed. Pgk1 was used as load control.</p
