Figure 2

CD63 localization in breast cancer cells. A) MDA-MB-231-luc-D3H2LN (MM231-LN) cells were transiently transfected with control (N.C.), RPN2 or CD63 siRNAs. After 2 days in culture, caspase-3/7 activity was assessed using an Apo-ONE Homogeneous Caspase-3/7 Assay system (left panel). The right panels show a representative image of Hoechst 33258-stained sections after N.C., RPN2 or CD63 siRNA transfection. The size bar indicates 50 μm. B) MCF7-ADR cells were transiently transfected with control (N.C.), RPN2 or CD63 siRNAs. After 2 days in culture, caspase-3/7 activity was assessed using an Apo-ONE Homogeneous Caspase-3/7 Assay system (left panel). The right panels show a representative image of Hoechst 33258-stained sections after N.C., RPN2 or CD63 siRNA transfection. The size bar indicates 50 μm. C) MM231-LN (upper panel) and MCF7-ADR cells (lower panel) were transiently transfected with N.C., RPN2 or CD63 siRNAs. After 1–3 days in culture, cell growth was assessed by an MTS assay. D) MM231-LN cells were grown and transiently transfected with N.C. (left panels) or RPN2 siRNA (right panels). After 2 days in culture, immunofluorescence of CD63 (Green) was detected. The blue color indicates nuclei. Cell membranes were stained PKH26 (Red). A higher magnification view is shown (CD63 HM). The size bar indicates 20 μm. E) MCF7-ADR cells were grown and transiently transfected with N.C. (left panels) or RPN2 siRNA (right panels). After 2 days in culture, immunofluorescence of CD63 (green) was detected. The blue color indicates nuclei. Cell membranes were stained PKH26 (red). A higher magnification view is shown (CD63 HM). The size bar indicates 20 μm. All data are shown as the mean  S.D., *P<0.05, **P<0.01.