Figure 5.

ERManI activity is required for ERAD substrate concentration in the ERQC. (A–C) NIH 3T3 cells transiently expressing H2a-RFP and ERManI-HA were pretreated without (A), with 10 μM lactacystin (B), or with 100 μM dMNJ (C) for 3 h before fixation and subjected to immunofluorescent staining of ERManI-HA with mouse anti HA-tag antibody and FITC-conjugated goat anti-mouse IgG. (D–F) H2aRFP was transiently coexpressed in NIH 3T3 cells with pSUPER-retro-GFP (expressing GFP from a PGK promoter, D and E) or the same vector encoding anti-ERManI shRNA in addition to GFP (F). Bar, 10 μm.