Dynamin inhibition affects secretion and degradation of pre-labeled [35S]-apoE.

HMDM were incubated in methionine/cysteine free DMEM with 250 µCi/mL [35S]methionine/cysteine for 3 h and received a preincubation with or without 30 µM MiTMAB during the last 30 min. Cells were then washed and chased in medium containing unlabelled methionine/cysteine, without or with 30 µM MiTMAB. At the indicated times [35S]-labelled apoE was immunoprecipitated from media and cell lysates, separated by SDS-PAGE and quantified by phosphorimaging. Secreted [35S]-apoE (A); cell-associated [35S]-apoE (B); net degradation of [35S]-apoE (C), calculated by subtracting residual [35S]-apoE in cell and media from [35S]-apoE at T0. Inserts show representative images. Fitted data according to Eq1 are shown (D–F). Symbols Ctrl (○) and MiTMAB (•) represent experimental data. *P