The TAD motif is essential for the TF function of Zas1. (A) Overview of mutant versions of zas1 cDNA alleles. Numbers indicate the amino acid residue range that has been deleted, replaced, or mutated in each construct. (B) Tetrad analysis of heterozygous zas1^c/zas1⁺, zas1^c-Δ276–282/zas1⁺, zas1^c-V276K, F280K/zas1⁺, zas1^c-Δ212–254/zas1⁺, zas1^c-Δ98–261/zas1⁺, and zas1^c-linker/zas1⁺ diploid fission yeast strains (C4083, C4098, C4740, C4387, C4114, C4486) after 5 d at 25°C. Circles identify the position of spores bearing the zas1^c, zas1^c-Δ276–282, zas1^c-V276K, F280K, zas1^c-Δ212–254, zas1^c-Δ98–261, and zas1^c-linker genes, respectively. (C) Comparison of Cnd1 protein levels in whole-cell extracts of asynchronous wild-type zas1⁺ and zas1^c-Δ276–282 strains expressing Cnd1 from the cnd1 or from the cnd3 promoter. Immunoblotting against α-tubulin serves as loading control. (D) Comparison of Cnd1 protein levels in asynchronous klf1⁺ and Δklf1 cells (C3974, C4573). Immunoblotting against α-tubulin serves as loading control. Comparison of condensation curves of wild-type (black, data from Fig. 5 E) and Δklf1 (red, C5141, n = 161 cells). (E) Tetrad analysis of SDD4/Δsdd4 S. cerevisiae diploid cells (C4743) after 2 d at 30°C. Comparison of Ycs4-PK₆ expression levels by immunoblotting against the PK epitope of whole-cell extracts of SDD4 and Δsdd4 cells derived from the tetrad dissection.