Ab106151

Approximately 40 mg of brain tissue lysed in RIPA buffer including protease inhibitors were homogenized and centrifuged at 12,000 rpm for 5 min at 4 °C. The concentration of protein was detected by a BCA Protein Quantitative Kit (Dingguo Changsheng Biotechnology, Beijing, China). Equivalent amounts of protein samples (40 ng) were separated by 10% SDS-PAGE and transferred to polyvinylidene fluoride membranes (PVDF). The antibodies used were as follows: APP 1:1000 (Abcam, ab126732, Cambridge, UK), SAA 1:1000 (Abcam, ab199030, Cambridge, UK), CYP27A1 1:1000 (Abcam, ab126785, Cambridge, UK), CYP7B1 1:1000 (ABclonal, A17872, Wuhan, China), CYP46A1 1:2000 (Abcam, ab244241, Cambridge, UK), RORγt 1:2000 (Abcam, ab207082, Cambridge, UK), Foxp3 1:1000 (Abcam, ab215206, Cambridge, UK), IL-17A 1:3000 (Abcam, ab189377, Cambridge, UK), GM-CSF 1:1000 (Proteintech, 17762-1-AP, Chicago, IL, USA), MIP-3α 1:1000 (Abcam, ab106151, Cambridge, UK), IL-10 1:1000 (Abcam, ab189392, Cambridge, UK), IFN-λ2 0.1 µg/mL (R and D, AF4635, Minneapolis, MN, USA). The protein density was measured by Image System Fusion FX (Vilber Lourmat, Paris, France) and GAPDH was used as the reference for standardization.

Approximately 40 mg brain or liver tissue were lysed in RIPA buffer including protease inhibitors. Then the tissues were homogenized and centrifuged (12,000 rpm, 5 min, 4 ℃) to collect the supernatant. The concentration of protein was measured by bicinchoninic acid (BCA) method. 40 µg protein samples were separated by 12.5% SDS-PAGE and transferred to PVDF membranes. The antibodies used were as below: RORγt 1:2000 (Abcam, ab207082), Foxp3 1:1000 (Abcam, ab215206), IL-17A 1:3000 (Abcam, ab189377), GM-CSF 1:1000 (Proteintech, 17762-1-AP), MIP-3α 1:1000 (Abcam, ab106151), IL-10 1:1000 (Abcam, ab189392), TGF-β1 1:1000 (Abcam, ab179695), IFN-λ2 0.1 µg/mL (R&D, AF4635), CYP27A1 1:1000 (Abcam, ab126785), CYP7B1 1:1000 (ABclonal, A17872), APP 1:1000 (Abcam, ab126732), and SAA 1:1000 (Abcam, ab199030). Image System Fusion FX (Vilber Lourmat, Paris, France) was used to detect the protein density.