Figure 4

Enhanced translocation and stronger activity of MIL5scFv-Arg9 to downregulate phospho-Akt in SKOV3 cells (HER2 positive). A: Confocal microscopy observation of MIL5scFv or MIL5scFv-Arg9 to penetrate SKOV3 cells; Cells were incubated with PBS (Control), MIL5scFv-Arg9-FITC (1:10) or MIL5scFv-FITC (1:10) at 37°C and collected consecutively in 0.5 h, 2 h and 5 h. The distribution of fluorescence signal in the cells was observed under a laser scanning confocal microscopy. Fluorescent signal could be detected after 0.5 hour’ treatment with MIL5scFv-Arg9 contrasting to MIL5scFv treated samples. Each panel was merged with two photos: FITC conjugated protein (green) and nuclei (blue, cells were stained with Hoechst 33258), which were not shown here; B: Effect of MIL5scFv or MIL5scFv-Arg9 on the expression of phospho-Akt. SKOV3 cells were treated with diluted MIL5scFv or MIL5scFv-Arg9 (4, 20 or 50 μg/ml, respectively) as indicated, and herceptin treated cells were set as positive control; C: Densities of phosphor-Akt were normalized based on densities of Akt shown in Figure4B, suggesting satisfactory enhanced effect of Arg9 in inhibiting the expression level of phospho-Akt in SKOV3 cells. The data shown are calculated from two independent experiments.