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. 2020 Jun 22;16(6):e1008905. doi: 10.1371/journal.pgen.1008905

Fig 6. Requirements of Pch2 binding in mitosis and meiosis.

Fig 6

A. Schematic of allele used for galactose-inducible mitotic expression of 3XHA-Pch2-E399Q. B. Western blot analysis of meiotic (i.e. endogenous) expression during meiotic G2/prophase at 4 hours after induction into the meiotic program and ectopic expression of pGAL10-3XHA-pch2-E399Q. Hours of treatment with galactose are indicated. C. ChIP-qPCR analysis of 3XHA-Pch2-E399Q at PPR1 (primer pair: GV2390/GV2391), GAP1 (primer pair: GV2597/GV2598) and HOP1 (primer pair: GV2605/GV2606) during in meiosis and mitosis. Time (hours) is indicated. Error bars represent standard error of the mean of at least three biologically independent experiments performed in triplicate. D. ChIP-qPCR analysis of active transcription (α−Rpo21-phospho-Ser5) GAP1 (primer pair: GV2597/GV2598), HOP1 (primer pair: GV2605/GV2606) in meiosis and mitosis. Hours are indicated. Error bars represent standard error of the mean of at least three biologically independent experiments performed in triplicate. E. ChIP-qPCR analysis of 3XFLAG-Pch2-E399Q at PPR1 (primer pair: GV2390/GV2391) and GAP1 (primer pair: GV2597/GV2598) in wild type or zip1Δ cells during meiotic G2/prophase (t = 4 hours). Error bars represent standard error of the mean of at least three biologically independent experiments performed in triplicate. F. Model depicting the interplay between Pch2 binding, active transcription, Orc1 and chromosome organization.