Cd9 20597 1 ap

C57BL/6 male mice (20–25 g, 8–10 wk old) were provided by the Animal Experiment Central at Chongqing Medical University (Chongqing, China). The ethical approval number of our institute is:2022-k276. HucMSCs were obtained from the Zhong Qiao Xin Zhou Biotechnology Co., Ltd. (Shanghai, China), and the human renal tubular epithelial cell line HK-2 was purchased from the Procell Life Science&Technology Co., Ltd. (Wuhan, China). antimycin A was purchased from Maokang Biotechnology Co., Ltd. (Shanghai, China), 2-Deoxy-D-glucose was purchased from Sigma Aldrich (St. Louis, MO, USA), Fetal bovine serum (FBS), PKH26, and PKH67 were purchased from Millipore Sigma (Burlington, MA, USA). The pAb anti-cleaved caspase-3 (9664S), Bcl-2 (3498), GRP78 (3177), IRE1-α (3294), XBP-1S (83418), CHOP (2895), p-PERK (3179), ATF6 (65880), and β-actin (3700) were obtained from Cell Signaling Technology (Danvers, MA, USA). The pAb anti-PDK4 (YN5701) was obtained from ImmunoWay Biotechnology Company (Plano, TX, USA). The pAb anti-TSG101 (28283-1-AP) and CD9 (20597-1-AP) were obtained from Proteintech Group, Inc. (Wuhan, China). The mimic miRNAs, inhibitor miRNAs, and small interfering RNAs (siRNAs) were purchased from Tsingke (Beijing, China). Lipofectamine 2000 was purchased from Thermo Fisher Scientific (Waltham, MA, USA).

Exosomes or cells were lysed with RIPA buffer containing a complete protease inhibitor tablet (Roche, Switzerland). Lysate was separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis, and the gel was blotted onto polyvinylidene fluoride (PVDF) membrane (Millipore, USA). The membrane was blocked in 5% nonfatmilk, and then incubated with either rabbit anti-human E-cadherin (3195, Cell Signaling Technologies, USA), N-cadherin (14,215, Cell Signaling Technologies, USA), Vimentin (5741, Cell Signaling Technologies), Twist (ab49254, Abcam, USA), PTEN (ab32199, Abcam, USA), CD103 (GTX64393, Genetex, USA), CD9 (20597–1-AP; Proteintech, USA), or Actin (3700, Cell Signaling Technology, USA). After washing, the membrane was incubated with the fluorescence-conjugated anti-mouse or anti-rabbit IgG (Invitrogen, USA). The bound secondary antibody was quantified using the Odyssey v1.2 software (LI-COR, USA) by measuring the band intensity (area × optical density) for each group and then normalized with Actin. The final results are expressed as fold changes by normalizing the data to control values.