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. 2018 Nov 15;27(1):188–199. doi: 10.1016/j.ymthe.2018.10.016

Figure 4.

Figure 4

Evaluation of Binding and Functional Activity of MdaPCSK9

(A) Western analysis of liver LDLR expression. Livers of control- and DMAb-treated C57B/6J wild-type mice were harvested at day 5 post-treatment. Levels of LDLR expression in tissue lysates were evaluated by staining for anti-mouse LDLR antibody. (B) Quantitative ELISA for MdaPCSK9 was performed on liver lysates at day 5 post-treatment. (C) Binding western blot analysis of anti-PCSK9 DMAbs from mouse sera. Anti-PCSK9 DMAbs obtained from mouse sera at 5 days post-treatment were evaluated for binding against recombinant mouse and human PCSK9 proteins. (D–F) Flow cytometry analysis was performed for LDLR expression in Huh7 cells. Human hepatoma Huh7 cells were treated with pVax-1 control DNA or MdaPCSK9 and harvested at 72 hr post-treatment. The level of LDLR expression was evaluated by flow cytometry by staining with anti-LDL receptor antibody (ab52818), followed by anti-rabbit IgG-FITC (sc-2012). (D) Histogram, (E) dot plot, and (F) mean fluorescence intensity (MFI) of Huh7 cells treated with pVax-1 or MdaPCSK9 are shown.