<p><b>A,</b> The TLS polymerases Eso1 and Kpa1 were upregulated in S phase. Eso1, Kpa1, and Rev7 protein expression levels were examined after <i>cdc25</i>-dependent block and release. Whole cell extracts were prepared by the boiling method, and western blotting was performed at different times (20–200 min) after the release. The graph represents the septation index after release. The lower panels show the expression patterns of Eso1, Kpa1, Rev7, Pcn1, Cdc13 (cyclin B), and Cdc2. <b>B,</b> Rev3 protein expression peaked during S phase. Whole cell extracts were prepared by the boiling method. The graph represents the septation index after release. The middle panel shows the expression pattern of Rev3, which was detected by anti-flag antibodies. The lower panel shows the protein expression of Pcn1 in the chromatin fraction. <b>C,</b> The protein amount of Rev1 was highest in G<sub>1</sub> phase. Whole cell extracts were prepared by the boiling method. The graph represents the septation index after release. The panels show the expression patterns of Rev1, Cdc13, and Cdc2. <b>D,</b> The protein expression of Rev1 in cell cycle mutants. <i>cdc10</i>, <i>cdc20</i>, <i>cdc22</i>, <i>cdc21</i>, <i>cdc17</i>, and <i>cdc25</i> mutants harboring flag-tagged <i>rev1</i> were arrested at 36.5°C for 4 h. Whole cell extracts were prepared by the boiling method. The amount of Rev1 was analyzed. <b>E,</b> Modified forms of the Rev1 protein were observed only in <i>cdc20</i> or <i>cdc22</i>. <i>cdc10</i>, <i>cdc20</i>, <i>cdc22</i>, and <i>cdc25</i> cells harboring flag-tagged <i>rev1</i> were arrested at 36.5°C for 4 h. Whole cell extracts were prepared by the LiNi method and then subjected to immunoprecipitation. Rev1 protein expression in each sample was roughly adjusted, and western blotting was performed.</p
