Fig. 2

Overexpression of ACSM5 inhibited FFA-induced lipid accumulation and fibrosis in LF cells. (A) CCK-8 assay was used to detect the effect of FFA on the proliferation of LF cells. (B) LF cells were treated with different concentrations of FFA for 48 h, and the mRNA and protein expression levels of ACSM5 in LF cells were detected by RT-qPCR and western blot. (C) ADV-ACSM5- or ADV-NC-infected LF cells were treated with FFA for 48 h, and the protein expression levels of ACSM5 in LF cells were detected by western blot. (D) ADV-ACSM5- or ADV-NC-infected LF cells were treated with FFA for 48 h, and oil red O staining was used to detect intracellular lipid accumulation in LF cells. Bar size = 100 nm. (E) ADV-ACSM5- or ADV-NC-infected LF cells were treated with FFA for 48 h, and TG assay kit was used to detect TG levels in LF cells. (F) ADV-ACSM5- or ADV-NC-infected LF cells were treated with FFA for 48 h, and the mRNA expression levels of FASN and ACC in LF cells were detected by RT-qPCR. (G and H) ADV-ACSM5- or ADV-NC-infected LF cells were treated with FFA for 48 h, and the protein expression of FASN and ACC in LF cells was detected by western blot. (G and I) ADV-ACSM5- or ADV-NC-infected LF cells were treated with FFA for 48 h, and the protein expression of collagen I, collagen III and α-SMA in LF cells was detected by western blot. LF, ligamentum flavum. FFA, free fatty acids. ADV, adenovirus. NC, negative control. ^n.s.P > 0.05, ^#P < 0.05, ^##P < 0.01, and ^###P < 0.001