Fig. 3: The role of Mnl1’s CTD in ERAD.
a, Mannosidase assays were performed with purified wild-type or mutant Mnl1–Pdi1 complex. b, Quantification of experiments as in a (see ‘Statistics and reproducibility’ in Methods). The means and s.d. of three experiments are shown. c, ERAD of CPY*–HA was determined by CHX chase experiments in cells lacking Mnl1 (mnl1Δ). The cells were transformed with wild-type Mnl1 or the indicated mutants. The samples were analyzed by SDS–PAGE and immunoblotting for HA. The intensities of the CPY*–HA bands were quantified. The fractions of CPY*–HA remaining at different time points are shown (means and s.d. of three experiments). d, Hydrophobic groove of the CTD; right, hydrophobicity scale. A semitransparent space-filling model in cartoon representation is shown. The three hydrophobic residues substituted in the mCTD mutant are labeled. e, Mannosidase assays were performed with the indicated purified wild-type or mutant Mnl1–Pdi1 complexes. f, Quantification of experiments as in e. The means and s.d. of three experiments are shown.
