Figure 1.

METTL3 (methyltransferase-like protein 3) increases PAI-1 (plasminogen activator inhibitor-1) expression and suppresses fibrinolysis in vitro. A, Human umbilical vein endothelial cells (HUVECs) were infected with control (scrambled) lentivirus or METTL3 shRNA lentivirus, and the mRNA level of METTL3 was analyzed by qPCR (n=3, data are mean±SEM). B, The protein level of METTL3 in shMETTL3 HUVECs was measured by Western blot (n=3, data are mean±SEM). C, Volcano plots showed differentially expressed transcripts in shMETTL3 HUVECs vs control HUVECs, including 183 downregulated genes and 132 upregulated genes (fold change >2 and adjusted P<0.05). D, Top gene ontology (GO) biological process categories enriched in downregulated and upregulated genes (blue color represents downregulated genes and red color represents upregulated genes). E, Heat map of top 20 differentially expressed genes in shMETTL3 HUVECs. F and G, The mRNA level (F) and protein level of PAI-1 (G) were determined in control and shMETTL3 HUVECs (n=3, data are mean±SEM). H, Fibrin formation and fibrinolysis were shown in shMETTL3 HUVECs and control cells (n=3, data are mean±SEM). I and J, The expression of PAI-1 was detected by Western blot (I) and quantitative real-time PCR (qPCR) (J) in METTL3 overexpressed and control cells (empty vector: EV, n=3, data are mean±SEM). K, Fibrin formation and fibrinolysis were shown in METTL3 overexpressed and control cells (n=3, data are mean±SEM). L and M, The expression of METTL3 and PAI-1 in shMETTL3 HUVECs infected WT METTL3 (shMETTL3+OE METTL3) and control cells (shMETTL3+EV) were measured by western blot (L) and qPCR (M) (n=3, data are mean±SEM). N, Fibrinolysis was shown in shMETTL3 HUVECs infected WT METTL3 (shMETTL3+OE METTL3) and control lentivirus (shMETTL3+EV) (Abs indicates absorbance, n=3, data are mean±SEM).