Anti cd36 apc

For in-vitro studies the following flow cytometry panel was used in conjunction with CTV and viability dye: anti-CD3 Alexa700, CD4 PE-Cy7, CD8 APC, CD44 PE all at a 1:200 dilution purchased from Biolegend. For in-vivo and ex-vivo studies, the following flow cytometry panel was used: anti-CD3 PE-Cy7, CD4 Pacific Blue, CD8 APC, CD44 PE, and anti-CD25 Alexa 700 all at a 1:200 dilution (except anti-CD25 at a 1:50 dilution) and were purchased from Biolegend. Endogenous Foxp3 expression was detected via eYFP fluorescence (or re-stained with anti-Foxp3 Efluor-450 clone FJK-16 s for any overnight staining (Fisher)), all acquisition was performed using a BD Fortessa flow cytometer. For surface FA transporter measurement, anti-CD36-APC (Biolegend), SLC27A1 (sigma), and SLC27A4 was purchased from Abcam (Cambridge, UK), followed by secondary goat anti-rabbit IgG Alexa-647 (Jackson Immunoresearch, West Grove, PA). Proliferation was analyzed using both FACS DIVA software as well as FlowJo software to determine expansion indexes.

Healthy Hispanics were enrolled in South Texas and evaluated for diabetes or dyslipidemias as described [4 (link), 19 (link), 20 (link)]. Mononuclear cells were isolated from heparinized blood, stored frozen (10% DMSO, 20% fetal bovine serum, 70% RPMI) and quick-thawed for batch phenotyping of baseline monocytes by flow cytometry. The antibodies and gating strategy to identify the three monocyte subpopulations (CD14+16- or classical, CD14+CD16+ or intermediate and CD14lo,CD16+ or non-classical) was described previously [14 (link)]. In addition, we used anti-HLA-DR-PE-Cy7 (eBioscience clone LN3) and anti-CD36-APC (Biolegend, clone 5–271). The MFI values were log-transformed for normalization and parametric analysis. Data analysis was conducted using SAS 9.4 with p values considered significant (≤ 0.5) or borderline significant (0.051 – 0.099).