Anti cdk1 sc 54

Cell extracts (30 μg) were prepared in RIPA lysis buffer (Santa Cruz Biotechnology, Santa Cruz, CA, USA) supplemented with protease and phosphatase inhibitor cocktail (Roche, Indianapolis, IN, USA) and separated on 12% SDS–PAGE gels. The expression levels of the indicated proteins were assessed by incubation with the following antibodies: anti-ID2 (NBP-88630; NovusBio), anti-TFCP2L1 (OAAB09732; Aviva Systems Biology, San Diego, CA, USA), anti-CDK1 (sc-54; Santa Cruz Biotechnology), anti-PARP (9542; Cell Signaling), anti-cleaved caspase-3 (9661; Cell Signaling), anti-β-actin (A5441; Sigma–Aldrich), and anti-Flag epitope (F3165; Sigma–Aldrich).

MSC-exos, cells and kidney tissues were lysed in RIPA lysis buffer (ThermoFisher) and the protein concentration was detected using BCA assay (ThermoFisher). Protein samples were subjected to Bis-Tris Gel (Invitrogen) and transferred onto PVDF membranes (Millipore). Membranes were blocked in NcmBlot blocking buffer (NCM Biotech) for 10 min and incubated overnight with primary antibodies as follows: anti-CD9 (ab92726, Abcam), anti-CD63 (sc-5275, Santa Cruz), anti-Tsg101 (sc-7964, Santa Cruz) anti-Alix (sc-53540, Santa Cruz), anti-GM130 (12480, Cell Signaling Technology), anti-CD44 (ab189524, Abcam), anti-ICAM-1 (MA5407, Invitrogen), anti-VCAM-1 (39036, Cell Signaling Technology), anti-LFA-1 (ab13219, Abcam), anti-integrin α4 (8440, Cell Signaling Technology), anti-integrin β1 (sc-374429, Santa Cruz), anti-PCNA (sc-56, Santa Cruz), anti-CDK1 (sc-54, Santa Cruz), anti-caspase-3 (ab13847, Abcam), anti-Cyclin B1(4138, Cell Signaling Technology), anti-p53 (2524, Cell Signaling Technology), anti-Bcl-2 (sc-7382, Santa Cruz), and anti-Bax (sc-20067, Santa Cruz). Secondary antibodies were used for detection by an ECL advanced system (GE Healthcare). Intensity values expressed as the relative protein expression were normalized to β-actin (AB2001, Abways) or GAPDH (AB2000, Abways).

Verapamil, vincristine, and paclitaxel were purchased from Sigma-Aldrich (St. Louis, MO, USA). The commercial antibodies used included anti-β-actin (sc-47778, dilution of 1:10,000), anti-CDK1 (sc-54, dilution of 1:1000), anti-cyclin B1 (sc-752, dilution of 1:1000), and α-tubulin (sc-23948, dilution of 1:5000) antibodies purchased from Santa Cruz Biotechnology (Dallas, TX, USA); anti-phospho-H3-S10 (#9701, dilution of 1:1000), anti-phospho-CDK1-Y15 (#4539, dilution of 1:1000), anti-CDK2 (#2546, dilution of 1:1000), anti-CDK4 (#12790, dilution of 1:1000), and anti-H3 (#9715, dilution of 1:5000) antibodies purchased from Cell Signaling Technology (Danvers, MA, USA); anti-INCENP (ab12183, dilution of 1:1000), anti-survivin (ab76424, dilution of 1:1000), anti-aurora B (ab2254, dilution of 1:1000), and anti-phospho-H3-T3 (ab78351, dilution of 1:1000) antibodies purchased from Abcam (Cambridge, UK); and anti-borealin (NBP1-89951, dilution of 1:1000) antibody purchased from Novus Biologicals (Centennial, CO, USA). HRP-conjugated secondary antibodies (dilution of 1:10,000) were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA).