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. 2021 Sep 8;297(4):101177. doi: 10.1016/j.jbc.2021.101177

Figure 2.

Figure 2

Comparative effects of PSCK9 on cellular LDLR and ASGR1 levels. HepG2-PCSK9-KO cells (A) or HepG2-naïve cells (B) were incubated for 20 h with media only (−PCSK9; SFM) or media containing purified PCSK9 (3 μg/ml) (+PCSK9). A, immunofluorescence microscopy of plasma membrane LDLR (green signal) and ASGR1 (red signal). The colocalization appears as yellow. The scale bar represents 15 μm. B, Western blot analysis of cellular LDLR and ASGR1 expression and of PCSK9 in the media of HepG2-naïve cells transfected with 20 nM nontargeting siRNA (siCnt). C and D, HepG2-naïve cells were transfected with nontargeting siRNA (siCnt) or siRNA PCSK9 (siPCSK9) at final concentrations of 10 nM and analyzed 48 h post transfection. C, quantitative RT-PCR analysis. D, Western blot analysis of cellular LDLR and ASGR1 and of secreted PCSK9. Data are representative of at least three independent experiments. Quantifications are averages ±SD. ∗p < 0.05; ∗∗∗p < 0.001 (t test).