Fig. 7.

Comparison of different amounts of MAP conjugation for MAP-polyplex cellular uptake and GFP silencing effect. MAP-polyplexes were prepared at with different concentrations of MAP conjugation. (a) To measure siRNA uptake, stably transfected Huh7.5 cells were treated for 6 h in OptiMEM. After treatment, cells were washed and lysates were collected. Cell lysates were measured at Ex. 544 and Em. 590 nm and normalized with protein concentrations quantified using BCA assay. A: No Treatment; B: 1.2 µM MAP; C: 50 nM siDY547; D: 50 nM siDY547-polyplex; E: 50 nM siDY547-polyplex + 1.2 µM MAP; F: 50 nM 0.5:1 CPP/PDP MAP-siDY547-polyplex; G: 50 nM 1:1 CPP/PDP MAP-siDY547-polyplex. (b) To measure GFP silencing, after a 6 h transfection and an additional incubation with Huh7.5 complete medium for 42 h, cells were washed and lysates were collected. Cell lysates were measured at Ex. 485 nm and Em. 518 nm and normalized with protein concentrations quantified using BCA assay. Error bars represent standard deviation (n = 3). A: 1.2 µM MAP; B: 50 nM siGFP; C: 50 nM siGFP-polyplex; D: 50 nM siGFP-polyplex + 1.2 µM MAP (unconjugated); E: 50 nM 0.5:1 CPP/PDP MAP-siGFP-polyplex; F: 50 nM 1:1 CPP/PDP MAP-siGFP-polyplex. Data are presented relative control treatments (A: Relative to non-treated cells; B–F: Relative to siNC equivalent counterpart). Asterisk (*) indicates p<0.05 based on analysis of variance and the Bonferroni test. Error bars represent standard deviation (n = 3).