Western blot analysis
Total protein extracted from kidney tissue was loaded into 10–12% SDS-PAGE gels and then transferred to polyvinylidene difluoride (PVDF) membranes (0.2 μm, Bio-Rad Laboratories, Inc.), as previously described. Immunoblots were visualized using and quantified by ImageJ software. Primary antibodies included: α-tubulin (Abcam, ab179484, 1:1000, RRID: AB_2890906), GAPDH (Zen BioScience, 200306-7E4, 1:5000, RRID: AB_2722713), collagen-I (Abcam, ab270993, 1:1000), collagen-IV (Abcam, ab6586, 1:1000, RRID: AB_305584), fibronectin (Boster Biological Technology, ba1772, 1:1000), α-SMA (Abcam, ab7817, 1:1000, RRID: AB_262054), TGF-β (Abcam, ab92486, 1:1000, RRID: AB_10562492), IL-1β (Abcam, ab9722, 1:1000, RRID: AB_308765), IL-6 (Huabio, EM170414, 1:1000), TNF-α (Huabio, R1203-1, 1:1000), ITGAM (Abcam, ab8878, 1:1000, RRID: AB_306831), SRC (Huabio, ET1602-03, 1:1000), p-SRC (Huabio, ET1609-15, 1:1000), FAK (Huabio, ET1602-25, 1:1000), p-FAK (Huabio, RT1216, 1:1000), Akt1 (Huabio, ET1609-47, 1:1000), p-Akt1 (Huabio, ET1701-36, 1:1000), GSK-3β (Huabio, ET1607-71, 1:1000), p-GSK-3β (Huabio, ET1607-54, 1:1000), β-catenin (Huabio, EM0306, 1:1000). The density quantitative analysis was conducted by Image J software.