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. 1998 Jul;9(7):1833–1845. doi: 10.1091/mbc.9.7.1833

Figure 5.

Figure 5

Cdc19p-containing MCM complexes do not change during the cell cycle. Each strain was shifted to 36°C for 4 h before harvest and lysis. Lanes 1: cdc10-V50 (G1 phase arrest, FY562); lanes 2: cdc22-M45 (early S phase, FY583); lanes 3: cdc17-K42 (late S phase, FY322); lanes 4: cdc25–22 (G2/M phase, FY584); lanes 5: wild-type asynchronous culture (FY254); lane 6: cdc19-P1 (S phase, FY243); lane 7: cdc21-M68 (S phase, FY786). (A) Equal amounts of cell lysate (10 μg total protein) were immunoblotted with antibodies to fission yeast MCMs (7% SDS-polyacrylamide gels) or α-tubulin (12% SDS-polyacrylamide gel). (B) Identical amounts of lysate (250 μg total protein) were immunoprecipitated with anti-Cdc19p antibody. Immunoprecipitates were washed with lysis buffer and immunoblotted for fission yeast MCM proteins, as indicated. (C) Flow cytometry measuring DNA content (X-axis) versus cell number (Y-axis) was used to verify cell cycle arrest.