Cyclin T2 (Ccnt2) and cyclin K (Ccnk) deficiency induces CHOP expression and calcification, opposite of cyclin T1 deficiency.
A, immunoblot analysis of CDK9, cyclin T1, cyclin T2, cyclin K, ATF4, CHOP, and GAPDH proteins in BSA (Veh) or 500 μm C18:0 treated scrambled (Scramb.) sg or Ccnt2 KO VSMCs for 6 h. B, qRT-PCR analysis of CHOP in BSA (Veh) or 250 μm C18:0 treated Scramb. sg or Ccnt2 KO VSMCs (n = 4). C, immunoblot analysis of CDK9, cyclin T1, cyclin T2, cyclin K, ATF4, CHOP, and GAPDH proteins in BSA (Veh) or 500 μm C18:0 treated Scramb. sg or Ccnk heterozygous KO VSMCs for 6 h. D, qRT-PCR analysis of CHOP in BSA (Veh) or 250 μm C18:0 treated Scramb. sg or Ccnk heterozygous KO VSMCs (n = 4). E, mineralization of Ccnt2 KO VSMCs. VSMCs were incubated with 2.0 mm phosphate with or without 250 μm C18:0 for 7 days (n = 6). F, mineralization of Ccnk hetero-KO VSMCs. VSMCs were incubated with 2.0 mm phosphate with or without 250 μm C18:0 for 7 days (n = 6). One-way ANOVA with a Student-Newman post hoc test was used for statistical analysis. *, p < 0.05.