Figure 1

Uptake of free-FAs and accumulation of TGs in FAs-treated cells. Amounts of Free-FAs (A) and TGs (B) were evaluated by thin-layer chromatography in cellular extracts of primary hepatocytes cultured with FAs mixture (2 : 1 oleate:palmitate with 1% BSA) to final concentration of 1 mM for the indicated times. Quantification of lipid spots was performed by scanning densitometry. The bars represent the means ± SD of the values, relatively to the corresponding control treatment, which was normalized to 100. n = 4; Comparisons were performed using Tukey’s honestly significant differences (HSD) test. *P < .05 vs control cells cultured without FAs; †P < .05 vs cells cultured with FAs for 1 hour; ‡ P < .05 vs cells cultured with FAs for 3 hours. (C) A representative silica gel plate separation of cellular lipid extractions is shown, with indication of TGs and free-FAs spots position. (D) Primary hepatocytes were cultured and treated with FAs mixture (2 : 1 oleate:palmitate with 1% BSA) or with FAs-Br mixture (2 : 1 oleate:2-Bromopalmitate with 1% BSA) to final concentration of 1 mM FAs for the indicated times. After that, cells were stained with Nile-Red and fluorescence was measured by FACS. *P < .05 vs control cells cultured without FAs; †P < .05 vs cells cultured with FAs for 1 hour; § P < .05 vs cells cultured with FAs-Br for 18 hours. (E) After the indicated treatments, Nile-Red stained cultures were examined under fluorescence microscopy (magnification × 400). Yellow fluorescence indicates intracellular accumulation of TGs.