Figure 2d.
(C,D) Localization of DTX in MD (C) or ID (D) cells. Cells were treated with MG132 (d–f) or leupeptin and NH4Cl (g–i) for 2 h, then fixed and permeabilized. DTX and LAMP-1 were detected using CY3–VSV and monoclonal 1D4B with an Alexa 488-coupled secondary antibody. The right panels are merges of the two adjacent panels, showing nuclei staining with Hoechst. Insets represent enlarged views (fivefold) of the boxed regions. DTX colocalizes with LAMP-1 in MD cells (white arrows in C) and is adjacent to LAMP-1 puncta in ID cells (yellow arrows in (D)). ID, DTX-transduced Itch−/− lines; MD, DTX-transduced wild-type lines; WB, western blot.