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. Author manuscript; available in PMC: 2012 Jun 1.

Published in final edited form as: Gastroenterology. 2011 Mar 9;140(7):1934–1942. doi: 10.1053/j.gastro.2011.02.063

KLB rs17618244 alters protein stability. Cycloheximide protein stability assay was performed using transient transfection into HEK293 cells with vectors expressing Myc-tagged KLB rs17618244 major allele G (KLB Arg728) or minor allele A (KLB Gln728). Cycloheximide (CHX, 20 μg/ml) was added to inhibit protein synthesis for a total of 0, 2, 4, 6, and 10 hours. Panel A shows representative Western blot analysis using anti-Myc. Anti-actin was used to verify equal protein loading. Panel B shows exponential best-fit curves generated from signal intensity measurements by densitometry of three independent experiments. Myc-KLB:actin ratios were calculated and normalized to time 0.

KLB rs17618244 alters protein stability. Cycloheximide protein stability assay was performed using transient transfection into HEK293 cells with vectors expressing Myc-tagged KLB rs17618244 major allele G (KLB Arg728) or minor allele A (KLB Gln728). Cycloheximide (CHX, 20 μg/ml) was added to inhibit protein synthesis for a total of 0, 2, 4, 6, and 10 hours. Panel A shows representative Western blot analysis using anti-Myc. Anti-actin was used to verify equal protein loading. Panel B shows exponential best-fit curves generated from signal intensity measurements by densitometry of three independent experiments. Myc-KLB:actin ratios were calculated and normalized to time 0.