The J-Domain of Heat Shock Protein 40 Can Enhance the Transduction Efficiency of Arginine-Rich Cell-Penetrating Peptides

Figure 3

Transduction of PTD-DsRed, DPV3A-DsRed, PTD-J-DsRed, and DPV3A-J-DsRed recombinant proteins. (a) 1.5 × 10⁵ Huh-7 cells in a well of 24-well plate were incubated with 0.2 mL of PTD-DsRed, DPV3A-DsRed, PTD-J-DsRed, or DPV3A-J-DsRed recombinant protein (40 μg/mL) for 0.5, 1, 2, 4, or 6 h. The cells were washed with PBS twice and the recombinant proteins incorporated into cells were released by 0.2 mL of 1% Triton X-100/PBS. After centrifugation to remove cell debris, 0.1 mL of the supernatant was transferred to a well of a 96-well plate and the amounts of recombinant proteins were measured by a fluorometer. The stimulating wave length and emission wave length were set at 557 nm and 585 nm, respectively. (b) 1.5 × 10⁵ Huh-7 cells in a well of 24-well plate were incubated with 0.2 mL of 5, 10, 20, and 40 μg/mL of the PTD-DsRed, DPV3A-DsRed, PTD-J-DsRed, or DPV3A-J-DsRed recombinant protein for 2 hours. The amounts of recombinant proteins were measured as described before. Experiments were done for four times and the average values were shown. All of the standard deviations were less than 1000. (c) The PTD-DsRed, DPV3A-DsRed, PTD-J-DsRed, and DPV3A-J-DsRed recombinant proteins were diluted in Huh-7 cell lysate to 0.05, 0.5, 2, and 5 μg/mL and the fluorescence values were measured as described above.

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