Fig. 8. Effects of ZF1/ZF2 mutations in the chromosomal ZAP1 gene on the regulation of gene expression. (A) Total RNA was extracted from exponential-phase cultures of the zap1 mutant strain ZHY6 (Δ) grown in LZM media supplemented with 3000 µM Zn²⁺ (lane 1) and from the wild-type strain, DY1457 and a chromosomal ZAP1^{ZF1&2 C2Q2} mutant, grown in LZM media supplemented with 3, 10, 30, 100, 300, 1000 and 3000 µM Zn²⁺ (lanes 2–8, respectively). The levels of ZRT1 mRNA were compared to the loading control CMD1 mRNA using S1 nuclease protection assays. Arrows indicate ZRT1 and CMD1 S1 nuclease protection products. (B) The band intensities in (A) were quantified and are plotted as the ratio of ZRT1:CMD1 mRNA levels at each zinc concentration. A representative of three independent experiments is shown.