Abca1

Immunoreactive bands were visualized using HRP-conjugated secondary antibodies and subsequent ECL detection (GE Healthcare Bio-Sciences Corp., Piscataway, NJ). Mouse or rabbit monoclonal antibody for ROCK II (Upstate, Lake Placid, NY), RhoA (Santa Cruz Biotechnology, Santa Cruz, CA), HMG-CoA reductase (Upstate), PPARγ (Abcam, Cambridge, UK), ABCA1 (Cell Signaling, Boston, MA), ApoA-I (Abcam), LXRα (Santa Cruz Biotechnology), LDLR (Abcam), HSL (Cell Signaling), eNOS (Abcam), and the loading control protein β-actin (Sigma-Adrich) were used in our Western blot analyses. Rabbit polyclonal antibody was used to recognize both PPARγ1 and PPARγ2 in experiments. Rp-8-pCPT-cGMPS and C3 exoenzyme were purchased from Sigma-Adrich. HFD was a basal purified diet (W/60% energy from fat, Blue:58G9 Test Diet; Richmond, VA). LDL was purchased from Abcam. Oxidized LDL (oxLDL) was purchased from Biomedical Technologies Inc. (Stoughton, MA).

Proteins were extracted from homogenized kidney tissues using RIPA buffer (Biosesang, Seongnam, Korea) and protein concentrations were determined using the BCA assay (Thermo Fisher Scientific, Waltham, MA, USA). Equal amounts of proteins were electrophoresed with glycine-sodium dodecyl sulfate buffer (LPS Solution, Daejeon, Korea) and transferred to polyvinylidene difluoride membranes (Millipore, Bedford, MA, USA) on ice. After the membranes were blocked with 5% skim milk (Becton Dickinson Rowa France, Le Pont-de-Claix, France) containing 2% bovine serum albumin, they were incubated overnight at 4° C under shaking with primary antibodies against E-cadherin (Abcam, Cambridge, UK), fibronectin, collagen, ABCA1, SREBP1, and GAPDH (Cell Signaling Technology). After 24 h, the membranes were incubated with a mouse or rabbit IgG-conjugated secondary antibody (Cell Signaling Technology) for 1 h. The protein bands were observed using the enhanced chemiluminescence method (Advansta, San Jose, CA, USA) and were quantified by densitometry using in ImageJ (National Institutes of Health, Bethesda, MD, USA).

Antibodies for Arg1 (Cat#: ab212522), CD31 (Cat#: ab28364) and ABCG1 (Cat#: ab52617) were purchased from Abcam (Cambridge, MA). Antibodies for IL-1β (Cat#: sc-52012), ICAM-1 (Cat#: sc-107), VCAM-1 (Cat#: sc-13160), ALP (Cat#: sc-365765), Οsx (Cat#: sc-393325) and RUNX2 (Cat#: sc-390351) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Antibodies for Cleaved Caspase1 (Cat#: 4199), ABCA1 (Cat#: 96292), β-actin (Cat#:4970) were purchased from Cell Signaling Technology (Danvers, MA). TG assay kit (Cat#:100020090), total cholesterol assay kit (Cat#: 100020080), LDL-C assay kit (Cat#:100020245), HDL-C assay kit (Cat#:100020235) were purchased from Biosino Bio-Technology and Science INC (Beijing, China). Alizarin Red S solution (Cat#: G3280) was purchased from Solarbio & Technology Co., Ltd (Beijing, China). IL-1β Elisa kit (Cat#: SEA563Mu), IL-6 Elisa kit (Cat#: SEA079Mu), TNF-α Elisa kit (Cat#: SEA133Mu), IL-10 Elisa kit (Cat#: SEA056Mu) and SOD Elisa kit (Cat#: SES134Mu) were purchased from Cloud-Clone Corp. (Wuhan, China). Mouse ROS Elisa kit (Cat#: YX-181519M) and Mouse ox-LDL Elisa kit (Cat#: YX-152412M) were purchased from Sino Best Biological Technology CO., Ltd (Shanghai, China). Calcium Colorimetric Assay kit (Cat#: MAK022-1KT) was purchased from Sigma-Aldrich. All other reagents were purchased from Sigma-Aldrich except where indicated.