<p><b>A</b>, the two-step marker switching method used for integration of the mutations. The genomic locus of <i>rad4<sup>+</sup></i> is shown on top. <b>B</b>, sensitivities of the cells with integrated <i>rad4</i> mutations to HU or MMS were examined by spot assay. The newly identified (upper part) and the previously reported (lower part) mutants are marked on the right. <b>C</b>, expression of Rad4 in the cells with the integrated <i>rad4</i> mutations was detected in whole cell lysates by Western blotting using anti-Rad4 antibodies. The asterisk denotes the cross-reactive material. Note: like the T45M mutant, A60P is a <i>ts</i> mutant.</p
