Figure 2.

Ab-MF3 inhibits PTS1-protein import in vitro. CHO cells were cultivated on coverslips, permeabilized with streptolysin-O, and incubated with HSA-SKL in transport buffer under various conditions, essentially as described (21). After washing and fixing, the coverslips were further processed by indirect immunofluorescence of HSA-SKL and quantitated as described in Materials and Methods. The results are expressed as the percentage of cells with peroxisomal HSA-SKL compared with the control standard. (1) + 75 molar excess CRYHLKPLQLKS (n = 2), (2) + 75 molar excess CRYHLKPLQSKL (n = 3), (3) − ATP-regenerating mix (n = 3), (4) + preimmune anti-HsPex5p (n = 2), (5) + anti-HsPex5p (n = 3), (6) + anti-hsp70 (n = 1), (7) + preimmune ab-MF3 (n = 3), and (8) + ab-MF3 (n = 3). (n refers to the number of separate experiments in which at least 250 cells in this condition were analyzed.)