FIGURE 8.

ALG-2·Sec31-PRR causes ablation of p24 by proteolytic degradation in an acidic compartment. Cells were transfected as indicated and then incubated for 8 h in regular medium or medium containing 100 μm leupeptin + 20 μm pepstatin or containing 100 μm chloroquine prior to fixation and immunofluorescence microscopy. At least 50 cells from each experimental condition were imaged, and each cell's p24 was scored (conditions were blinded, and images were randomly interspersed) as either normal, partially disrupted, or ablated. Data shown are from a single experiment but are qualitatively similar to several other experiments obtained under similar conditions. The condition for the fifth group of bars (no RFP) employed a wild type untagged ALG-2 construct, demonstrating that the p24 phenotype was not dependent upon the RFP tag.