Acat1

Aliquots of protein extracts (20 μg) derived from THP1 monocytes, their derived macrophages, HUVECs, and HASMCs were separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis, and then immunoblotted with specific antibodies raised against the following proteins: CD68, ACAT1, ICAM1, VCAM1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), ABCA1, collagen 1 (Novus Biologicals, Littleton, CO, USA), collagen 3, fibronectin, arginase 1, phospho (Ser529)-NFκB, α-tubulin, MMP2 (GeneTex, Irvine, CA, USA), MMP9 (EnoGene, Atlanta, GA, USA), elastin, MARCO, selectin E (Bioss, Woburn, MA, USA), PPARγ (Signalway Antibody, College Park, MD, USA), phospho (Thr202/Tyr204)-ERK1/2, phospho (Ser/Thr)-Akt (Cell Signaling Technology, Tokyo, Japan), c-Src (Bioworld Technology, St. Louis Park, MN, USA), PI3K, Bcl2, Bax, and β-actin (Sigma). The band intensity of the immunoblot was quantified by densitometry [39 (link),40 (link),41 (link),44 (link),45 (link),46 (link),47 (link),48 (link),49 (link)].

Aliquots of 20 µg of protein extracts from human macrophages and HASMCs were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and subjected to immunoblotting with the following antibodies: CD36 (R&D Systems, Minneapolis, MN), ACAT1 (Santa Cruz Biotechnology, Santa Cruz, CA), ABCA1, collagen-1 (Novus Biologicals, Littleton, CO), collagen-3, fibronectin, α-tubulin, MMP2 (GeneTex, Irvine, CA), MMP9 (EnoGene, Atlanta, GA), elastin (Bioss, Woburn, MA), or β-actin (Sigma) [16] (link)–[19] (link). The densities of the bands were measured using a Densitograph System (Ez-Capture II and CS Analyzer 3.0, ATTO, Tokyo, Japan).

Antibody against LDLR (Cat # ab-30532),  MCT-1 (Cat # ab-90582), MCT-3 (Cat # ab-60333) and SR-B1 (Cat # 217318) were purchased from Abcam (Cambridge, UK); antibodies against FASN (Cat # sc-55580), MCT-4 (Cat # sc-50329), MCT-2 (Cat # sc-166925), LDHA (Cat # sc-137244), LXRα/β (Cat # sc-377260), ACAT-1 (Cat # sc-517387), ACAT-2 (Cat # sc-293307), phospho-ERK (Tyr 204) (Cat # sc-7383), ERK (Cat # sc-154), Cyclin A (Cat # sc-751), CDK 2 (Cat # sc-163), Cyclin E (Cat # sc-198), β-tubulin (Cat # sc-9104), GAPDH (Cat # sc-20357), β-actin (Cat # sc-1615), and HRP conjugated secondary antibodies: anti-rabbit (Cat # sc-2030), anti-mouse (Cat # sc-2031), anti-goat (Cat # sc-2033) were purchased from Santa Cruz Biotechnology (CA, USA). Human LDL cholesterol (Cat # 360–10) and human HDL cholesterol (Cat # 361–10) were purchased from Lee Biosolution (MO, USA); Water soluble-free cholesterol and Dextran sodium sulfate (DSS) (Cat # 160110) were purchased from MP Biomedicals (CA, USA). Azoxymethane (AOM) (Cat # A5486), 2-Deoxy-D-glucose (2-DG) (Cat # D8375) and Oxamate (Cat # O2751) were purchased from Sigma Aldrich (MO, USA).