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. 2019 Dec 10;20(24):6220. doi: 10.3390/ijms20246220

Figure 5.

Figure 5

BFR1 deletion does not affect PMT1 and PMT2 transcription. (A) RT-PCR analysis of PMT1 and PMT2 mRNA levels in wild type (BY4741) and bfr1Δ strains. Cells were grown in YPD medium, total RNA was extracted and cDNA was prepared and used as a template for RT-PCR. Results show mRNA abundance with respect to TAF10 mRNA from three independent experiments ± SD. For statistical significance a multiple t-test was performed. (B) Western blot analysis of Pmt2 protein levels expressed under the control of a GAL1 inducible promotor in total cell extracts from strains described in (A) upon addition of indicated sugars. (C) RT-PCR analysis of PMT2 mRNA levels in strains described in (A) in which Pmt2 is expressed under the control of a GAL1 inducible promotor upon growth in galactose containing media. Results show mRNA abundance with respect to ACT1 mRNA from three independent experiments ± SD. For statistical significance an unpaired t-test was performed. (D) Cycloheximide chase analysis of Pmt1 and Pmt2 in wild type (BY4741) and bfr1Δ strains. Cycloheximide was added to a final concentration of 100 to 200 µM and equal amounts of cells were sampled at indicated time points and subjected to Western blot analysis using Pmt1 and Pmt2 antibodies. G6PDH served as a loading control. N.s. = not significant.