Fig. 2.

CIP1 expression at M/G1 is mediated by Mcm1 through ECB elements on CIP1 promoter. a After synchronized cells had been released from G1 phase, total RNA was extracted from WT and ecb I  +  ecb II strains and analyzed by Northern blotting. b The quantitative results of the relative amount of CIP1 normalized to the internal control, ACT1, was shown. c Schematic diagram indicating two potential ECB elements on the CIP1 promoter. The mutation sites that destruct two ECB elements are shown below. d Reporter analysis was used to determine the contribution of ECB elements on the promoter activity of CIP1. The CIP1 promoter was placed at the upstream of the lacZ gene and then integrated into the yeast genome. To create the ecb I  +  II mutant, the ECB elements on the integrating plasmid were mutated before insertion. The lacZ expression was normalized to ACT1. The values were given as mean ± s.d. (n = 3,**P < 0.01, Student’s t-test, two tailed). e ChIP analysis of Mcm1 binding at CIP1 promoter and the times indicated the minutes after 0.5 M KCl treatment. The binding of Mcm1 on CIP1 promoter was normalized with U2 background. The values were given as mean ± s.d. (n = 3, *P < 0.05, **P < 0.01, Student’s t-test, two-tailed)