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. 2010 Oct 18;5(10):e13488. doi: 10.1371/journal.pone.0013488

Figure 4. Loss of Hip4 impairs transcriptional silencing.

Figure 4

(A) Centromeric (dh-dg) repeat silencing. Strains containing the otrR1::ade6+ allele in combination with the appropriate deletions were grown to log phase in YE5S medium subjected to 5-fold serial dilutions and spotted on to YE5S agar lacking adenine. Plates were incubated for 3–4 days at 30°C. (B) Centromeric transcripts accumulate in HIRA mutants. RNA purified from the indicated strains was subjected to strand-specific RT-PCR analysis. One primer complementary to the cenFOR or cenREV transcript was included for the reverse transcription step and the second primer was added during the PCR amplification step. Control reactions omitting the reverse transcription step (-RT) were included to demonstrate the absence of DNA. (C) Strains containing the mat3-M::ade6+ allele in combination with the appropriate deletions were treated as described for (A). Experiments were performed at least twice and a representative example is shown.