Dapi staining solution c1006

A human mesangial cell line (HMC cell line, FH0241) was obtained from FuHeng Biology Co. (Shanghai, China). DHT (116064-77-8, purity ≥ 98%) was acquired from Krre Technology Co. (Beijing, China). Mannitol (69-65-8) was obtained from Macklin Biochemical Co. (Shanghai, China). The following antibodies: GAPDH (1:1000, ab8245), α-smooth muscle actin (1:1000, ab7817), Collagen I (1:1000, ab260043), Fibronectin (1:1000, ab2413), PI3KCA (1:1000, ab40776), PI3K (1:1000, ab191606), p-PI3K (1:1000, ab182651), AKT (1:1000, ab179463), and p-AKT (1:1000, ab192623) were bought from Abcam Co. (Cambridge, United Kingdom). Goat Anti-Rabbit IgG H&L (HRP, A0208), Rabbit Anti-Mouse IgG H&L (HRP, A0216), and Rabbit Anti-Mouse IgG H&L (Alexa Fluor 488, A0428) were purchased from Beyotime (Shanghai, China). DAPI Staining Solution (C1006) and Cell Counting Kit-8 (C0038) were also obtained from Beyotime (Shanghai, China). Glucose, Trypsin, Dulbecco’s Modified Eagle’s Medium (DMEM), and fetal bovine serum (FBS) were purchased from GIBCO. The Mouse Albumin ELISA Kit (ab207620) was purchased from Abcam.

Phospholipid (soybean lecithin containing 90.4% phosphatidylcholine) was purchased from Shanghai Tai-wei Pharmaceutical Co. Ltd. (Shanghai, China). Fusidic acid was obtained from Shanghai Aladdin Biochemical Technology Co. Ltd. (Shanghai, China). Trifluoroacetic acid (TFA, analytical grade) and Coumarin 6 (C6) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Luria–Bertani Medium was supplied by Coolaber Science & Technology Co., Ltd. (Beijing, China). DAPI staining solution (C1006) was purchased from Beyotime Biotech Inc. (Shanghai, China).

Tetramethylpyrazine (TMP) was purchased from Yuanye Bio-Technology .Co, Ltd (Shanghai, China). LPS from Escherichia coli 0111:B4 (LPS) and Dexamethasone (Dex) was obtained from Sigma-Aldrich (St.Louis, MO, United States). Primers for qRT-PCR were synthesized in Tsingke Biological Technology .Co, Ltd (Beijing, China). Primer details were listed in Quantitative Real-Time PCR Assay. Antibodies used for Western blot included GAPDH (ab8245), NFκB p65 (ab16502), NFκB P-p65 (ab194726), TRAF6 (ab33915), caspase-1 (ab74279), caspase-1 (ab 1872), caspase-3 (ab90437) from Abcam (Cambridge, United Kingdom), and β-actin (3700s), ASC (67824T), NLRP3 (1510s), caspase-8 (4927T) from Cell Signal Technology (Danvers, MA, United States), TLR4 (66,350-1-lg) from proteintech (Rosemont, IL, United States). Anti-rabbit (7074s) and anti-mouse (7076s) secondary antibodies conjugated with HRP were from Cell Signal Technology (Danvers, MA, United States). Antibodies used for Immunofluorescence contained iNOS (18985-1-AP), CD206 (18704-1-AP) and CD68 (66231-2-Ig) were purchased from Proteintech Group (Rosemont, IL, United States). Secondary antibodies conjugated with FITC (a0568), and DAPI staining solution (c1006) were from Beyotime (Shanghai, China).