The expression of <i>cis4</i>, <i>zrg17</i>, and <i>zhf1</i> is not dependent upon zinc.

<p><b>(A)</b> Total RNA was isolated from wild-type and the indicated mutants grown overnight in ZL-EMM supplemented with 0 or 100 μM Zn²⁺. The levels of <i>cis4</i>, <i>zrg17</i>, and <i>zhf1</i> transcripts were compared to the loading control <i>act1</i> using S1 nuclease analysis. A representative experiment is shown in the left panel and the average values from three independent experiments are shown on the right, with error bars representing standard deviations. ND = Not determined. (B). S1 nuclease analysis was performed as described above with the exception that mRNA was hybridized to probes complementary to <i>zrt1</i> and <i>act1</i>. Radiolabelled <i>act1</i> probes were diluted by 10-fold with unlabeled probe for experiments with <i>cis4</i>, <i>zrg17</i>, and <i>zhf1</i>. When cells were grown under zinc-limiting conditions, lower levels of <i>zrt1</i> transcript accumulated in the <i>cis4</i>Δ and <i>zrg17</i>Δ mutants compared to the wild-type. <i>p</i> values were determined using a student's t-test.