Figure 3.

Protein gel electrophoresis (chip). (A) Pro260 chip automated gel electrophoresis. Five controls and five high-fat-diet left ventricle protein gel electrophoresis obtained from Ossabaw swine heart. Basically, 4 μL of denatured protein for each sample was loaded into a Pro260 Experion chip (Bio-Rad). (B) Experion Pro260 Ladder electropherogram. The Pro260 ladder contains nine proteins ranging from 10–260 kDa as well as 1.2 kDa lower marker. (C and D) Sample electropherograms obtained from control (C) and high-fat-fed (D) left ventricle protein extractions. Left ventricles were homogenized in radio immunoprecipitation assay lysis buffer (which included phenylmethylsulfonyl fluoride and Na orthovanadate solutions as well as protease inhibitor cocktail). The homogenates were centrifuged at 45,000 rpm for 30 minutes at 4°C. The supernatants were collected and used for analysis.