FIGURE 2.

CsA inhibits apoE secretion and degradation in HMDM. HMDM were incubated in methionine/cysteine-free Dulbecco's modified Eagle's medium with 250 μCi/ml [³⁵S]methionine/cysteine for 3 h and received a preincubation of 10 μm CsA during the last 1 h. Cells were then washed and chased in medium containing unlabeled methionine/cysteine, without (○) or with (●) 10 μm CsA. At indicated times ³⁵S-labeled apoE was immunoprecipitated from media and cell lysates, separated by SDS-PAGE, and quantified by phosphorimaging as described under “Experimental Procedures.” A, secreted ³⁵S-labeled apoE; B, cell-associated ³⁵S-labeled apoE; C, net degradation of ³⁵S-labeled apoE (calculated by subtracting residual ³⁵S-labeled apoE in cell and media from ³⁵S-labeled apoE in cells at T₀). Data are expressed as a percentage of the total ³⁵S-labeled apoE present at T₀. Symbols represent actual experimental data, which were analyzed by two-way repeated measures analysis of variance, analyzing for effect of time and treatment. A, p < 0.01 for control versus CsA in medium; B, p < 0.01 for control versus CsA in cell; C, p = not significant for control versus CsA net degradation. Lines show fitted data according to the Equation 1.