Author: Labrie, Marilyne; Lalonde, Simon; Najyb, Ouafa; Thiery, Maxime; Daneault, Caroline; Des Rosiers, Chrisitne; Rassart, Eric; Mounier, Catherine
Title: Apolipoprotein D Transgenic Mice Develop Hepatic Steatosis through Activation of PPAR? and Fatty Acid Uptake Document date: 2015_6_17
ID: 0wtq1c15_22
Snippet: Primary hepatocytes from WT and Tg mice were cultured in serum free media for 48h. Fatty acid uptake was measured using 3 H-oleate as previously described [47] . Briefly, cells were incubated in 0.68 μCi/mL 3 H-oleate (50 μM)_bound to BSA (fatty acid/BSA molar ratio 2:1) in serum free DMEM/F12 media for 10 min at RT. The reaction was stopped by adding 200 μM of ice-cold phloretin solution for 2 min. Cells were then washed 3 times with PBS and .....
Document: Primary hepatocytes from WT and Tg mice were cultured in serum free media for 48h. Fatty acid uptake was measured using 3 H-oleate as previously described [47] . Briefly, cells were incubated in 0.68 μCi/mL 3 H-oleate (50 μM)_bound to BSA (fatty acid/BSA molar ratio 2:1) in serum free DMEM/F12 media for 10 min at RT. The reaction was stopped by adding 200 μM of ice-cold phloretin solution for 2 min. Cells were then washed 3 times with PBS and lysed in 0.1 N NaOH for 30 min at RT. Radioactivity in lysates was counted in 10 mL Ultima-Gold solution (Tri-Carb 2800TR, Perkin Elmer) and protein were quantified (Bradford Assay, BioRAD).
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