Figure 5

In contrast to aggresomes, dispersed aggregates of CyPrP^EGFP do not affect the localization of poly(A)⁺-RNA or protein translation. (A) N2a and Hela cells were transiently transfected with CyPrP^EGFP. After 24 h, cells were fixed, permeabilized, and processed for in situ hybridization to detect poly(A)⁺ RNA (Red), and analysed by fluorescence microscopy. Asterisks indicate transfected cells. (B) Total protein production was measured by analyzing newly synthesized proteins in N2a and Hela cells transfected with empty vector, EGFP, or CyPrP^EGFP, as indicated. Cells were labelled with 25 μCi/ml ³⁵S labelling mix in methionine (Met)- and cysteine (Cys)-deficient medium. Where indicated, medium contained 30 μg/ml cycloheximide (CHX). Equal amounts of proteins were separated by 10% SDS-PAGE and radioactivity signals were determined with a phosphorimager. The position of the molecular mass markers is indicated on the right.