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. 2024 Mar 30;15:2789. doi: 10.1038/s41467-024-46336-2

Fig. 1. PCSK9 directly switched on pro-inflammatory genes and increased the expression of adhesion molecules.

Fig. 1

a Immunoblot analysis demonstrated that proprotein convertase subtilisin/kexin type-9 (PCSK9) activated and phosphorylated nuclear factor (NF)-κB p65 in THP-1 (N = 5) and human umbilical vein endothelial cells (HUVECs, N = 3) in a dose-dependent manner (0, 50, 200, and 2000 ng/mL). b Luciferase assay demonstrating that the NF-κB gene promoter was significantly activated (P < 0.001) after 12-h treatment with PCSK9 (200, 2000 ng/mL, N = 5), resistin (10, 50 ng/mL, N = 5) or TNF-α (10, 20 ng/mL, N = 3) in HEK293T cells. c qPCR analysis demonstrating that PCSK9 significantly increased the expression of cytokines, including TNF-α (N = 6), IL-1β (N = 5), IL-6 (N = 6) and IL-10 (N = 5), and integrin-α4 (N = 3) and -β1(N = 3) in a dose-dependent manner (0, 50, 200, and 2000 ng/mL) in monocytes. d qPCR analysis demonstrating that PCSK9 significantly increased the expression of cytokines, including TNF-α (N = 7), IL-1β (N = 6), and IL-6 (N = 7) and C-reactive protein (CRP, N = 7) in a dose-dependent manner (0, 50, 200, and 2000 ng/mL), while albumin (N = 7) was not increased in hepatocytes. e qPCR analysis demonstrated that PCSK9 significantly increased the expression of pro-inflammatory cytokines, including TNF-α (N = 6), IL-1β (N = 5), and IL-6 (N = 5), as well as adhesion molecules (VCAM-1 (N = 6), ICAM-1 (N = 6), E-selectin [SELE, N = 6]) in HUVECs. f Immunoblot analysis demonstrating that PCSK9 increased the protein levels of integrin-α4 and -β1 in THP-1 cells (N = 4) and VCAM-1 and ICAM-1 in HUVECs (N = 3) in a dose-dependent manner (0, 50, 200, and 2000 ng/mL). g Fluorescence-activated cell sorting analysis reveals a 16% increase in VLA-4 activation on monocyte surfaces with PCSK9 treatment, compared to 2.7% in the vehicle group. h Immunoblot analysis demonstrating that PCSK9 treatment (2 µg/mL) for 40 min activated and phosphorylated NF-κB in BMDMs from Ldlr−/− mice and BL6 control mice (N = 4). i qPCR analyses revealed PCSK9-induced cytokines (TNF-α, IL-1β, and IL-6) even in Ldlr−/− BMDMs (N = 5) after 24-h treatment (2 µg/mL). The differences between the groups were compared using the unpaired t-test (two-tailed). All experiments were independently performed, and data are presented as mean values ± SD.