Vitro 3.three. ADAR2 Prevents Hepatocyte from Lipid Accumulation In VitroTo investigate the effects of ADAR2 on hepatic lipogenesis, loss-of-function and To investigate the effects of ADAR2 on hepatic lipogenesis, loss-of-function and gain- gainof-function analyseswere performed on HepG2 cells. Initial, the efficiency ofoverexpressof-function analyses have been performed on HepG2 cells. Initial, the efficiency on the the overexpressing (OE) ADAR2 plasmid was confirmed. The ADAR2 level was significantly improved ing (OE) ADAR2 plasmid was confirmed. The ADAR2 level was substantially elevated afterafter transfection together with the ADAR2 overexpression plasmid in HepG2 cells (Figure We We transfection using the ADAR2 overexpression plasmid in HepG2 cells (Figure 3A). 3A). utilized siRNA to induce the silence of ADAR2, and si-ADAR2-2 was chosen among 3 made use of siRNA to induce the silence of ADAR2, and si-ADAR2-2 was chosen among three sesequences resulting from its higher knockdown efficiency (Figure 4A). The expression of ADAR2 quences due roughly two-fold reduced compared(Figure 4A). group. Nile Red staining mRNA was to its high knockdown efficiency to the control The expression of ADAR2 mRNA was that ADAR2 overexpression wascompared to the 3B) and group. Nile Red staining indicated approximately two-fold reduced lowered (Figure control that ADAR2 knockindicated that ADAR2 overexpression was decreased (Figure 3B) and vitro.ADAR2 knockdown (Figure 4B) accelerated the lipid droplet formation of HepG2 cells in that Furthermore, ADAR2 overexpression decreased the droplet formation of HepG2 cells in vitro. Furthermore, down (Figure 4B) accelerated the lipidlevels of TG (Figure 3C), although ADAR2 knockdown could overexpression of TG (Figure 4C). Meanwhile, PCR analysis showed that ADAR2 ADAR2 elevate the levels decreased the levels of TG (Figure 3C), although ADAR2 knockdown overexpression levels of TG (Figure 4C). Meanwhile, PCR ADAR2 knockdown procould elevate thedownregulated lipid formation (Figure 3D) andanalysis showed that ADAR2 moted lipogenesis (Figure 4D). These results give evidence that ADAR2 could possibly be a crucial regulator of lipogenesis through NAFLD.Nutrients 2023, 15, x FOR PEER REVIEW7 ofNutrients 2023, 15,overexpression downregulated lipid formation (Figure 3D) and ADAR2 knockdown promoted lipogenesis (Figure 4D).Nisin Z Autophagy These final results provide evidence that ADAR2 may well be a7cruof 17 cial regulator of lipogenesis through NAFLD.Sinensetin supplier Figure three.PMID:24120168 ADAR2 overexpression suppresses lipogenesis in hepatocytes. (A) The mRNA levels of Figure 3. ADAR2 overexpression suppresses lipogenesis in hepatocytes. (A) The mRNA levels of ADAR2 relative to GAPDH had been analyzed by qRT-PCR right after transfection with the ADAR2 overexADAR2 relative to GAPDH have been analyzed by qRT-PCR soon after transfection together with the ADAR2 overexpression plasmid in HepG2 cells. (B) The lipid content material of HepG2 cells was determined by Nile Red pression plasmid in HepG2 cells. (B) The lipid content of HepG2 cells was determined by Nile Red staining. (C) The TG levels of HepG2 cells. (D) The mRNA levels of lipogenesis-related genes (Fasn and Srebp1c) relative to GAPDH had been analyzed by qRT-PCR. p 0.001; n = 6. OE, overexpression; OA, oleic acid; TG, triglyceride; ADAR2, adenosine deaminases acting on RNA two; Fasn, fatty acid synthase; Srebp1c, sterol regulatory element binding protein-1c. Scale bar = one hundred .Nutrients 2023, 15, x FOR PEER REVIEW8 ofNutrients 2023, 15,staining. (C) The TG levels of HepG2 cells. (D) The mRNA levels of lipogenesis-re.
