Fig. 3.

Cyclin K is loaded onto chromatin in parallel with pre-RC. a Cyclin K protein level detected by immunoblotting in soluble (free) and chromatin-bound (CB) fractions with or without DNase treatment. b Loading of cyclin K and components of prereplicative complex (CDT1, MCM2, and MCM7) onto chromatin. HCT116 cells were synchronized at M phase (0 h), and then released into fresh medium. Corresponding cell cycle profiling by FACS analyses was presented in right panel. c Loading of cyclin K and Mcm2 onto chromatin. NIH 3T3 cells were synchronized at G0 by serum starvation, and then released into fresh serum-containing medium. Corresponding cell cycle profiling by FACS analyses was presented in the right panel. d Loading of cyclin K, MCM2, and MCM7 onto chromatin. HCT116 cells were synchronized at G1/S boundary by hydroxyurea treatment (HU, 1 mM), and then released into fresh medium. Corresponding cell cycle profiling by FACS analyses was presented in right panel. e Loading of cyclin K and Mcm2 onto chromatin during the course of liver regeneration in vivo. 0 h denotes samples collected immediately after partial hepatectomy. All experiments were repeated more than three times and representative results are shown