Ab218528

The protein used for WB is derived from the supernatant or cell lysate. Cell culture medium was concentrated by Amicon Ultra-4 Centrifugal Filter Devices (Amicon Ultra 50 K device, 50,000 MW CO, Millipore, USA) after centrifugation at 7500 × g for 40 min. The condensed samples (approximately 200 µl) were mixed with 5× loading buffer, boiled for 3 min and then analyzed by WB. Cell lysates obtained after treatment of cells with different inhibitors. AT2 cells were treated with PS1145 at 20 μM for 30 min, or JSH-23 (#S7351, Selleck Chemicals, China) at 6 μM for 1 h, or C646 at 20 μM for 1 h, or Anacardic Acid (#S7582, Selleck Chemicals, China) at 10 μM for 1 h, respectively. After 48 h, cell lysates were collected. The protein was separated on a sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to PVDF membranes (Millipore, USA), and blocked with 5% non-fat dry milk in TBST. After three times of washing with TBST, following primary antibodies dissolved in antibody buffer (Keygentec, China) were used: anti-ECM1 (sc-365335, SANTA CRUZ, USA), anti-p-p65 (S536) (#3033, CST, USA), anti-Ac-p65 (K310) (#3045, CST, USA), anti-t-p65 (#8242,CST, USA), anti-p300 (#86377, CST, USA), anti-integrinα6 (ab181551, Abcam, USA), anti-integrin β4 (ab236251, Abcam, USA), anti-ABCG1(ab218528, Abcam, USA), anti-HA (#3724, CST, USA), and anti-β-actin (#4970, CST, USA).

Primary rabbit polyclonal antibodies against AMPK (2532), p-AMPK (2535), mTOR (2983), p-mTOR (39182), ACC (3676), p-ACC (11818), SCD-1 (2794S), ATGL (2439S), CPT-1 (97361S), and goat anti-rabbit IgG (whole molecule) were purchased from Cell signaling technology, Inc. (Danvers, MA, USA). Primary rabbit polyclonal antibodies against ABCA1 (ab18180), ABCG1 (ab218528), LC3 (ab192890), SREBP-1c (ab28481), FAS (ab133619), and β-actin (ab8227) were purchased from Abcam (Cambridge, MA, USA). Primary rabbit polyclonal antibodies against α-SMA (14395), CD36 (18836), Beclin1 (11306), ATG5 (10181), and ATG7 (10088) were purchased from Proteintech Group, Inc. (Rosemont, IL, USA). TG and total cholesterol (TC) assay kits were purchased from BioSino Bio-Technology and Science Inc. (Beijing, China). T0 and MGF were purchased from MedChemExpress, Int. (Shanghai, China).