Validation of regulated genes by RT-PCR analysis of mRNA from polysome fractions. Total RNA was extracted from polysome fractions of wild type (wt) (left panels) and myo1Δ (right panels) strains. A) RT-PCR analysis of ribosomal protein mRNAs RPL7B, RPL3, RPS8A. B) RT-PCR analysis of mRNAs SRL3, PST1, PIR3, YPS4, and KTR2 regulated in response to cell wall damage. The RT-PCR products were resolved on 2% agarose gels, stained with ethidium bromide, and normalized against ACT1, a cytoskeletal protein mRNA that was not regulated in the microarray hybridization study. The lane numbers at the top of each gel correspond to the sucrose density gradient fractions shown in Figure 2. Below each set of gels is shown the results of densitometry analysis of RT-PCR products from duplicate experiments. See Methods for details.
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