Trilencer 27 human sirna duplexes

To determine cell surface expression of CD36, cells were labelled with anti-human CD36 antibody (FA6-152, Stem Cell Technologies, Vancouver, Canada) or isotype control (IgG 1 κ) at a concentration of 10 µg/ml in RPMI1640 media (Sigma-Aldrich, Merck, Sydney Australia) containing 10% FBS. Cells were then washed and stained with a secondary antibody, goat anti-mouse IgG H&L Dylight 650 (Clone ab96882, Abcam, Cambridge, United Kingdom) at a concentration of 5 µg/ml. Cell viability was determined via 7AAD (BD Biosciences) staining at a 1:20 dilution. Samples were processed using a BD Accuri C6 ow cytometer (BD, Becton, Dickinson and Company, New Jersey, USA) and data analyzed using FCS Express 4 Flow Research Edition (De Novo software, Los Angeles, USA).
CD36 knockdown using siRNA CD36 targeting small interfering RNAs (siRNAs) Trilencer-27 Human siRNA duplexes at 20µM were purchased (Origene, Maryland, USA) with three CD36 targeting siRNAs (A, B and C) plus a non-targeting scrambled (SCR) siRNA used as a negative control. Transfection was performed using Lipofectamine RNAiMAX (Invitrogen by Life Technologies, California, USA) as per the manufacturer's instructions with the knockdown maximized at 5nM and validated by ow cytometry.