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. 2006 Feb;8(2):125–135. doi: 10.1593/neo.05556

Figure 3.

Figure 3

Internalization of GrB/scFvMEL into A375-M, MEL-526, TXM-18, and TXM-1 cells, as assessed by immunofluorescent staining. Cells were treated with 40 nM GrB/scFvMEL for 1 hour. Molecules bound to the cell surface were removed by brief treatment with glycine buffer (pH 2.5). Cells were fixed in 3.7% formaldehyde and permeabilized in 0.2% Triton X-100. Samples were blocked with 3% BSA, incubated with goat anti-GrB mAb, and then incubated with FITC-coupled anti-goat IgG and PI. The slides were mounted with DABCO containing 1 µg/ml PI and analyzed under a Nikon Eclipse TS-100 fluorescence microscope. Internalization of GrB/scFvMEL was observed in the gp240 antigen -positive A375-M, MEL-526, and TXM-18 cells, but not in the gp240 antigen-negative TXM-1 cells. (A) No GrB/scFvMEL treatment control. (B) GrB/scFvMEL treatment for 1 hour.