FIG 5.
Cell cycle-dependent expression of NSun2 and CDK1. (A) U2OS cells were synchronized by serum starvation for 3 days or were left unsynchronized (Asyn.). Cells released from arrest by addition of 10% fetal bovine serum (FBS) at the times indicated were subjected to FACS analysis to monitor the cell cycle distribution. (B) Cell lysates prepared from the cells described for panel A were subjected to Western blot analysis to assess NSun2, CDK1, and GAPDH proteins. (C) RNA isolated from the cells described for panel A at times 0 and 18 h was subjected to bisulfate RNA sequencing to assess the methylation levels of endogenous CDK1 mRNA. Open boxes indicate cytosine-to-uracil conversion, read as thymidine levels in the cDNA (nonmethylated [Unmet.]), and filled boxes indicate a retained cytosine (methylated [Met.]). The numbers below the columns refer to cytosine positions in the CDK1 3′UTR.