CDK2 arrests AML cell differentiation by directly targeting and activating PRDX2. (A) The interaction between HA-CDK2 and PRDX1/2/6 was detected by immunoprecipitation. HL60 cells were transfected with CDK2-HA plasmid for 3 days. After immunoprecipitation with HA, their interaction were detected by western blotting with PRDX1/2/6 antibodies. (B) Western blotting of PRDX2/6 and CDK2 in shCDK2-transduced HL60 cells. (C) The effect of CDK2 on the peroxidase activities of PRDX2/6. The recombinant PRDX2/6 protein was incubated with recombinant CDK2 for 1 hour and peroxidase activity was monitored for 60 minutes. (D) Silencing efficiency of shRNA against PRDX2 in AML cells (HL60, U937, and NB4). (E-G) Cell differentiation analysis of AML cells infected with lentivirus-shPRDX2 for 7 days. (E) CD11b expression. (F) Cell morphological analysis after Wright-Giemsa staining. (G) NBT-reducing activity in HL60 cells. (H) Differentiation-related protein expression in AML cells infected with lentivirus-shPRDX2. The protein levels of STAT1, C/EBPβ, MAFB, and PRDX2 were measured by western blotting. (G) ***P < .001 vs scramble. Data are shown as mean ± SD, n = 3. Other data are representative of at least 3 individual experiments; 1 representative image is shown.