Fig. 7: CircSARS-CV2-N1368 regulates the phenotypes of HCMECs by activating TLR4/NF-κB/ROS signal. | Acta Pharmacologica Sinica

Fig. 7: CircSARS-CV2-N1368 regulates the phenotypes of HCMECs by activating TLR4/NF-κB/ROS signal.

From: CircSARS-CV2-N1368 from SARS-CoV-2 impairs endothelial cell function through the upregulation of ATF7 to activate TLR4/NF-κB/ROS signaling

Fig. 7Fig. 7

a Volcano plot showing the dysregulated genes in HCMECs by circSARS-CV2-N1368 and ATF7 siRNA, respectively. CircSARS-CV2-N1368-up-regulated genes and ATF7 siRNA-down-regulated genes in HCMECs were overlapped in Venn diagram (b) and categorized by KEGG enrichment analysis (c). d, e The expression of Toll-like receptor-related genes in HCMECs with overexpression of circSARS-CV2-N1368 or knock-down of ATF7 by RT-qPCR assay. f Transcriptional activation of TLR4 gene by dual luciferase assay. g Detection of ATF7, TLR4 and NF-κB p65 in circSARS-CV2-N1368 overexpressing HCMECs with immunofluorescence assay. h Expression of ATF7, TLR4 and NF-κB p65 activation in HCMECs by Western blot assay. i VEGFA expression, activations of eNOS, HH3 and NF-κB p65 in HCMECs by Western blot assay. j Detection of ROS level in HCMECs by using DCFH-DA probe. k Proliferation activity of HCMECs by EdU assay. l, m Migration activity of HCMECs by trans-well migration assay and wound healing assay, respectively. n Matrigel tube formation assay of circSARS-CV2-N1368-overexpressiong HCMECs with knock-down of ATF7, TLR4 and NF-κB p65, respectively. Comparisons were made with unpaired t test in (d)–(f), and comparisons were made with 2-way ANOVA in (h), (i), and 1-way ANOVA in (j)–(n), n = 3 per group. The scale bar is 50 μm in (g), (j), and (l), the scale bar is 100 μm in (n), the scale bar is 200 μm in (m). *P < 0.05, **P < 0.01, ***P < 0.001.

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