Fig. 2: Silencing or functional inhibition of NPM in LX-2 cells significantly decreased the expression of liver fibrosis markers α-SMA, collagen I, and MMP9.

A Western blot results showed that when NPM in CSFC-2G and LX-2 cells was knocked down by CRISPR-Cas9, the expression levels of α-SMA, collagen I, and MMP9 were sharply downregulated. B Quantitative PCR confirmed the decreased mRNA expression of α-SMA, collagen I, and MMP9 in LX-2 and CSFG-2G cells with NPM knocked out. C Western blot analysis results showed that NPM knockdown by siRNAs decreased fibrosis markers, including α-SMA, collagen I, and MMP9 in LX-2 cells. D Quantitative PCR further confirmed the NPM knockdown by siRNAs and the reduced α-SMA, collagen I, and MMP9. E NPM protein was knocked down or overexpressed in primary hepatic stellate cells cultured for 13 days, and markers of liver fibrosis were decreased or increased correspondingly. F CIGB300, a small molecule inhibitor of Ser125 phosphorylation of NPM protein, downregulated α-SMA, collagen I, and MMP9 in LX-2 cells. The data shown are representative of three independent experiments and relative quantification of data based on β-actin or α-tubulin gene expression. n ≥ 3; mean ± SEM; **p < 0.01, ***p < 0.001, ****p < 0.0001; T test.