Computational protocol: Lipid Accumulation in HepG2 Cells Is Attenuated by Strawberry Extract through AMPK Activation

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Protocol publication

[…] Equal amounts of protein (75 µg) of each sample were loaded and separated in a 10% acrylamide sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS/PAGE) and transferred to nitrocellulose membranes (Bio-Rad) by using a Trans-Blot Turbo Transfer System (Bio-Rad Laboratories, Inc., Hercules, CA, USA). Membranes were washed three times with PBS, blocked with 5% non-fat dry blocker (Bio-Rad), and incubated with the primary antibody at 4 °C overnight. All the polyclonal antibodies for the protein of interest were purchased from Santa Cruz Biotechnology, Inc., Dallas, TX, USA. Next, membranes were washed with 5% Tween 20 in Tris HCl buffered saline and incubated with the secondary anti-rabbit immunoglobulin G-Peroxidase antibody (Sigma-Aldrich, Milan, Italy). Protein bands were visualized using Immobilon Western Chemiluminescent Substrate (Millipore Corporation, Billerica, MA, USA), and the protein signals were detected by a Lycor C-Digit Blot Scanner. Quantification of protein expression was made using the software (Image Studio 3.1) provided by the manufacturer of the Blot Scanner (LI-COR Biotechnology, Bad Homburg, Germany). [...] Statistical analyses were performed using STATISTICA software (Statsoft Inc., Tulsa, OK, USA). Data were subjected to one-way analysis of variance for mean comparison, and significant differences among different treatments were calculated according to Tukey’s HSD (honest significant difference) multiple range test. Data are reported as mean ± standard deviation (SD). Differences at p < 0.05 were considered statistically significant. All the analyses were performed in triplicate. […]

Pipeline specifications

Software tools Image Studio, Statistica
Applications Miscellaneous, DNA fingerprinting
Organisms Homo sapiens
Diseases Cardiovascular Diseases, Diabetes Mellitus, Carcinoma, Hepatocellular
Chemicals Acetyl Coenzyme A, Ampicillin, Cholesterol, Fatty Acids, Triglycerides