Fig 2. Sec61p N-acetylation at S2 is important for ERAD.

ERAD of CPY* was investigated in sec61S2Y (A), sec61ΔH1, and sec61ΔN21 (B). Cells were grown at 30°C to an OD₆₀₀ = 1, translation was inhibited by adding 200 μg/ml cycloheximide, extracts were prepared by bead-beating, and samples resolved by SDS-PAGE. The 0' samples were taken in duplicate in (B). Proteins were transferred to nitrocellulose and CPY* was detected with polyclonal rabbit antiserum and enhanced chemiluminescence. Bands were quantified using the ImageQant TL software. Sec61 (A) and Rpn12 (B) were used as loading controls. Band intensities were quantified relative to loading controls. Curves represent the averages of three independent experiments. Error bars represent standard deviations.