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. 2008 Oct 16;113(2):338–346. doi: 10.1182/blood-2008-08-172924

Figure 3.

Figure 3

Expression of human perforin in RBL cells. (A) Flow cytometry of RBL-1 cells expressing recombinant human perforin cDNA, wild-type (WT) or mutant, after retroviral transduction. The retroviral vector contains GFP in cis with the perforin cDNA. GFP expression is shown in the left column, perforin expression in the right column. The results of 3 independent transductions are summarized in the bar graph with the MFI of the WT protein set at 100% and the MFI of all mutant perforins compared with WT. *Statistical significance compared with WT (2-tailed Student t test, P < .05). (B) Western blot of WT and mutant perforins expressed in RBL-1 cells. Three different monoclonal antibodies were used (P1-8, 2d4, and Pf-344). The precursor and mature isoforms seen in the WT perforin are marked by arrows. The lysates were generated under nonreducing conditions. (C) Metabolic study: Western blot of lysates from perforin-expressing, RBL-1 cells treated overnight with an alkalinizing agent (CMA), a cysteine protease inhibitor (E64), or no treatment (control; −).