Figure 5.

(A) Representative immunoblot showing that in PC3-PIP cells the down-regulation of Chk following 24 h of incubation with nanoplex 1 was dependent on the concentration of siRNA-Chk incorporated into the nanoplex (N/P ratio is 50). GAPDH protein levels were used for protein loading assessment. Lane 1: PC3-PIP cells treated with nanoplex 1 without siRNA-Chk. Lane 2: PC3-PIP cells treated with nanoplex 1 with 100 nM scrambled siRNA. Lane 3: PC3-PIP cells treated with nanoplex 1 with 100 nM siRNA-Chk. Lane 4: PC3-PIP cells treated with nanoplex 1 with 50 nM siRNA-Chk. Lane 5: PC3-PIP cells treated with nanoplex 1 with 20 nM siRNA-Chk. (B) Therapeutic efficacy of siRNA and prodrug in PC3-PIP cells. PC3-PIP cells were treated with nanoplex 1 without siRNA-Chk (control), nanoplex 1 with siRNA-Chk (siRNA-Chk), nanoplex 1 without siRNA-Chk but with 5-FC (5-FC), and nanoplex 1 with siRNA-Chk and 5-FC for 24, 48, and 72 h (siRNA-Chk+5-FC). (1, Treatment for 24 h; 2, treatment for 48 h; 3, treatment for 72 h; nanoplex concentration = 350 nM, N/P = 50, siRNA-Chk concentration = 80 nM, 5-FC concentration = 3 mM. Values represent mean ± SEM of three or more assays for each treatment; *, P < 0.05; **, P < 0.01; ***, P < 0.001.)