Fig 8. The expression of cis4, zrg17, and zhf1 is not dependent upon zinc.

(A) 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 cis4, zrg17, and zhf1 transcripts were compared to the loading control act1 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 zrt1 and act1. Radiolabelled act1 probes were diluted by 10-fold with unlabeled probe for experiments with cis4, zrg17, and zhf1. When cells were grown under zinc-limiting conditions, lower levels of zrt1 transcript accumulated in the cis4Δ and zrg17Δ mutants compared to the wild-type. p values were determined using a student's t-test.