Journal: International Journal of Molecular Sciences
Article Title: Oncogenic H-Ras Expression Induces Fatty Acid Profile Changes in Human Fibroblasts and Extracellular Vesicles
Figure Lengend Snippet: Gene expression analysis of fatty acid-metabolizing enzymes in control and H-RasV12 expressing fibroblasts by qRT-PCR. ( A ) Gene expression analysis of desaturases ( SCD and FADS ), elongases ( ELOVL ) and acyl-coenzyme A synthetases ( ACSL ). ( B ) Gene expression analysis of phospholipases A2 ( PLA2 ). Ten ng of each cDNA were used as template. Reactions were performed in triplicate, using SYBR green technology StepOne RT-PCR machine to detect amplification. The GAPDH gene was used as endogenous control. The fold expression in H-RasV12 fibroblasts with respect to control is displayed, expressed as Relative Quantity (RQ). The analysis was repeated three times in triplicate and the mean ± S.D. is reported (* p
Article Snippet: 4.8. qRT-PCR RNA was extracted and retro-transcribed as previously described [ ]. cDNA was used to determine the expression of genes listed in . cDNA was used to determine transcript levels by qRT-PCR in a StepOne RT-PCR machine (Applied Biosystems, Foster City, CA, USA) using SYBR® Select Master Mix (Life Technologies, Carlsbad, CA, USA).
Techniques: Expressing, Quantitative RT-PCR, SYBR Green Assay, Step One RT-PCR, Amplification