Cdk4 d9g3e

Compounds 1, 2, and 3 were synthesized according to the procedure developed by our group [12 (link)]. They were dissolved in dimethyl sulfoxide (DMSO), the concentration of which never exceeded 0.1% (v/v); 50 mM of stock solution was stored at −20 °C. The Cell counting Kit-8 (CCK-8), Hoechst 33258, JC-1, and the ROS assay kit were purchased from Beyotime Institute of Biotechnology Company (Shanghai, China). The annexin V-FITC and propidium iodide (PI) kit was obtained from BD Pharmingen (BD, San Diego, CA, USA). The primary antibodies used are as follows: CDK4 (D9G3E, Cell Signaling Technology, Danvers, MA, USA), Cyclin D1 (E3P5S, Cell Signaling Technology), Caspase-3 and cleaved Caspase-3 (D3R6Y, Cell Signaling Technology), PARP (46D11, Cell Signaling Technology), Cytochrome c (AF0146, Affinity, Changzhou, China), Caspase-9 (AF6348, Affinity), cleaved Caspase-9 (D8I9E, Cell Signaling Technology), BAX (D2E11, Cell Signaling Technology), Bcl-2 (D17C4, Cell Signaling Technology), Bad (D24A9, Cell Signaling Technology), and β-actin (AF0003, Beyotime Biotechnology, Shanghai, China). All other chemicals used are commercially available and reagent grade.

The following antibodies were used: ß-actin (Sigma-Aldrich, A5441, mouse antibody, 1:5000 dilution), glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Cell Signaling Technology, 5174s, rabbit antibody, 1:5000 dilution), RBM39 (Atlas Antibodies, HPA-001591, rabbit antibody, 1:2000 dilution), c-Myc (Cell Signaling Technology, 5605s, rabbit antibody, 1:2000 dilution), MYCN (Santa Cruz Biotechnology, 53993, mouse antibody, 1:200 dilution), FLAG (Sigma-Aldrich, F1804, mouse antibody, 1:2000 dilution), ATM (D2E2) (Cell Signaling Technology, 2873, rabbit antibody, 1:1000 dilution), EZH2 (D2C9) (Cell Signaling Technology, 5246, rabbit antibody, 1:1000 dilution), CDK4 (D9G3E) (Cell Signaling Technology, 12790, rabbit antibody, 1:1000 dilution), RET (C31B4) (Cell Signaling Technology, 3223, rabbit antibody, 1:1000 dilution), and EED (Millipore 09-774, rabbit antibody, 1:1000 dilution).