Figure 2.

In Vitro Evaluation of MdaPCSK9

(A) In vitro expression of MdaPCSK9 compared to empty backbone pVax-1 plasmid in HEK293 cells. Supernatants were harvested at 24, 48, and 72 hr post-transfection and analyzed by quantitative ELISA. (B) Binding western blot analysis of MdaPCSK9 from cellular supernatants. Binding of MdaPCSK9 obtained from HEK293T transfected cells was evaluated against recombinant mouse and human PCSK9 proteins. Membranes were stained with supernatant from MdaPCSK9- or pVax-1-transfected cells at 72 hr post-transfection. (C) Immunofluorescence staining of mouse Hepa1-6 cells transfected with MdaPCSK9 plasmids. Cells were fixed 48 hr after transfection. Cells were stained with anti-mouse IgG-FITC and DAPI nuclear stain. pVax-1-transfected cells were used as a negative control.