Pparγ

SDS-PAGE and western blots were performed using standard protocols. Antibody binding to bands was detected using an ECL detection system (Sage Creation). The primary antibodies used were specific to MUC17 (Sigma-Aldrich, St Louis, MO, USA), IL-1β (Bioworld, St Louis Park, MN, USA), IL-8 (Bioworld), cdx1 (Abcam), pJNK (Affinity biosciences, Cincinnati, OH, USA), pERK (Santa Cruz, Dallas, TX, USA), p38 (Cell Signaling Technology, Danvers, MA, USA), pp38 (Cell Signaling Technology), PPARγ (Affinity biosciences), pp65 (ZSGB-Bio, Beijing, China), p65 (ZSGB-Bio), p21^waf (Proteintech, Rosemont, IL, USA), p53 (ZSGB-Bio), Myh9 (Abcam, Cambridge, UK), and Actin (Sigma-Aldrich).

After treatment with PMFs and ox-LDL for 24 h, RAW264.7 cells were washed with PBS twice and lysed with RIPA on ice for 30 min. The protein concentration was determined by the BCA protein assay kit (Beyotime Biotechnology, Shanghai, China). In total, 40 μg proteins were separated on 8% SDS-PAGE gels and transferred to the polyvinylidene difluoride (PVDF) membrane (250 mA, 90 min). Then, the blots were blocked with 5% bovine serum protein for 1.5 h and incubated with the antibodies overnight at 4°C. The following antibodies were used: SRA1 (1:1,000), CD36 (1:1,000), ABCG1 (1:1,000), PPARγ (1:1,000), and LXRα (1:1,000) were purchased from Affinity Biosciences (Jiangsu, China). SRB1 (1:2,000) was purchased from Abcam (Cambridge, England). GAPDH (1:1,000) was purchased from Cell Signaling Technology Inc. (Boston, MA, United States). Followed by the appropriate HRP-conjugated rabbit anti-rabbit IgG (1:5,000, Cell Signaling Technology Inc., Boston, MA, United States) for 1 h, visualized by the enhanced chemiluminescence (ECL) reagent and exposed to Gel Doc XR+ (Bio-Rad, Hercules, CA, United States). The bands density was analyzed and calculated using the Image Lab software.

Antibodies from Affinity Biosciences: p-Smad1/5/8 (1:1000, #AF8313), Smad1/5/8 (1:1000, #AF0614), PPARγ (1:1000, #bs-4590R), UCP1 (1:1000, #72298). Antibodies from Beyotime: HA tag (1:1000, #AH158), Flag tag (1:1000, #AF519). Antibodies from Sangon Biotech: p-ACC (1:1000, #D155180), ACC (1:1000, #D155300). Antibodies from Abcam: BMP8A (1:1000, #154373). Antibodies from ZenBio: p-AMPK (1:1000, #R26252), AMPK (1:1000, #380431), PGC-1α (1:1000, #381615). Antibodies from CWBIO: goat anti-Rabbit IgG HRP secondary antibody (1:4000, #CW0103S), goat anti-Mouse IgG HRP Conjugated (1:4000, # CW0102). Antibodies from Bioss: p38 MAPK (1:1000, #bs-0637R), p-p38 MAPK (1:1000, #bs-2210R), β-actin (1:2000, #bs-0061R), p-JNK (1:1000, #bs-1640R), JNK (1:1000, #bs-2592R), C/EBPα (1:1000, #AF6333), p-Smad2/3 (1:1000, #AF3367), Smad2/3 (1:1000, #AF6367), p-ERK1/2 (1:1000, #AF1015), ERK1/2 (1:1000, #AF0155), p-p65 (1:1000, #AF2006), p65 (1:1000, #AF5006), p-IKKα/β (1:1000, #AF3013), IKKα/β (1:1000, # AF6014).