Fig. 2: HBx upregulates NLRP3-mediated pyroptosis and MASH-associated METTL3 in HBx-expressing hepatocytes in vitro. | Cell Death & Disease

Fig. 2: HBx upregulates NLRP3-mediated pyroptosis and MASH-associated METTL3 in HBx-expressing hepatocytes in vitro.

From: Hepatic steatosis and pyroptosis are induced by the hepatitis B virus X protein via B56α-METTL3 interaction-mediated m6A modification of the NLRP3 mRNA

Fig. 2

A–L HepG2 cells were transfected with pcDNA3.1-HBx (1 μg/ml, 24 h) to construct HBx-expressing hepatocytes, while the negative control (NC) cells transfected with pcDNA3.1 vector served as controls. A, B Levels of TC (A) and TG (B) in cells. C Relative mRNA levels of NLRP3 and pyroptosis-related genes were measured by qRT-PCR. D Levels of HBx expression and NLRP3, ASC, Caspase-1, GSDMD, and IL-1β proteins were detected by WB. E The activity of caspase-1 related to pyroptosis was quantified by ELISA. F Representative images showing co-localization of GSDMD-N (red) and Dio (green) were captured by confocal microscopy (Left). Scale bar, 10 µm. Quantification of the relative intensity of GSDMD-N/Dio is shown in a bar graph (Right). G–I Levels of LDH (G), IL-1β (H), and IL-18 (I) in supernatants. J Global RNA m6A levels in cells were measured by colorimetric assays. K Relative mRNA levels of METTL3, METTL14, and WTAP were detected by qRT-PCR. L Expression levels of METTL3, METTL14, and WTAP proteins were detected by WB. (M-N) A liver transcriptome dataset (GSE89632) was derived from NASH patients (n = 19) and healthy controls (HC, n = 24). M Correlation analysis between methyltransferase-related genes and inflammation- or lipid metabolism-related genes. N GSEA was performed. Data are presented as mean ± SD. *, P < 0.05, compared to the control group.

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