ILF3 inhibits p-AMPK expression to drive non-alcoholic fatty liver disease progression

Figure 1 Elevated ILF3 expression in non-alcoholic fatty liver disease patients and models. A: ILF3 expression in serum of non-alcoholic fatty liver disease (NAFLD) patients; B: Serum ILF3 expression according to NAFLD severity; C and D: Elevated ILF3 expression in rising Oleic acid mixture (OA) concentration treated in HepG2 cells; E and F: Oil Red O staining showing increased lipid deposition in OA-treated HepG2 cells and mice liver tissues; G and H: Elevated ILF3 expression in NAFLD mice model detected by qPCR and Western blotting. All data are presented as the mean ± SD. ^bP < 0.01; ^cP < 0.001; NAFLD: Non-alcoholic fatty liver disease; SCD: Standard chow diet; HFD: High-fat diet.

Figure 2 Knockdown of ILF3 alleviated lipid deposition in hepatocyte and mice models. A and B: Knockdown of ILF3 reduced triglyceride (TG), aspartate transaminase (AST) and alanine transaminase (ALT) secretion in Oleic acid mixture (OA)-treated HepG2 cells; C: Oil Red O staining showing knockdown of ILF3 Lightens lipid accumulation in OA-treated HepG2 cells; D-F: Knockdown of ILF3 suppressed the proliferation and promoted the apoptosis of hepatocytes; G and H: Knockdown of ILF3 reduced TG, AST and ALT secretion in mice with high-fat diet (HFD) feeding; I: Oil Red O staining shows knockdown of ILF3, which lightens lipid accumulation in liver tissues with HFD feeding. All data are presented as the mean ± SD. ^bP < 0.01; ^cP < 0.001; AST: Aspartate transaminase; ALT: Alanine transaminase; NC: Negative control; shILF3: Small hairpin RNA targeting LF3 intravenously.

Figure 3 Knockdown of ILF3 alleviated non-alcoholic fatty liver disease progress by activating the AMPK pathway in vitro. A and B: Knockdown of ILF3 was helpful to phosphorylation of the AMPK pathway in Oleic acid mixture (OA)-treated HepG2 cells detected by Western blotting; C: Oil Red O staining showing knockdown of ILF3 decreased lipid deposition in OA-treated HepG2 cells, while CompoudC reversed this effect; D and E: Knockdown of ILF3 reduces triglyceride, alanine transaminase and aspartate transaminase secretion in OA-treated HepG2 cells, while CompoudC reversed this effect. All data are presented as the mean ± SD. ^aP < 0.05; ^cP < 0.001; AST: Aspartate transaminase; ALT: Alanine transaminase; NC: Negative control; shILF3: Small hairpin RNA targeting LF3 intravenously.

Figure 4 Knockdown of ILF3 alleviated non-alcoholic fatty liver disease progress by activating the AMPK pathway in vivo. A and B: Western blotting showing knockdown of ILF3 helped activate the AMPK pathway in liver tissues; C: Oil Red O staining showed knockdown of ILF3 decreased lipid deposition in liver tissues, while CompoudC reversed this effect; D and E: Knockdown of ILF3 reduced triglyceride, alanine transaminase and aspartate transaminase secretion in mice with high-fat diet feeding, while CompoudC reversed this effect; F: Knockdown of ILF3 was helpful to upregulate pAMPK expression, while CompoudC can reverse this effect. All data are presented as the mean ± SD. ^bP < 0.01; ^cP < 0.001; AST: Aspartate transaminase; ALT: Alanine transaminase; NC: Negative control; shILF3: Small hairpin RNA targeting LF3 intravenously.