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. Author manuscript; available in PMC: 2015 Apr 15.
Published in final edited form as: Science. 2015 Apr 3;348(6230):132–135. doi: 10.1126/science.1260638

Fig. 4. epe1 mutants retain heterochromatin without tethered Clr4 methyltransferase through multiple cell divisions and meiosis.

Fig. 4

(A) wild-type, epe1Δ, epe1-K314A and epe1-H297A cells carrying 4xtetO-ade6+ and expressing TetR-Clr4*, were grown −/+ AHT. Colony colour assay to assess 4xtetO-ade6+ silencing (red-pink colonies; % of total indicated) and H3K9me2 qChIP on 4xtetO-ade6+ with (−AHT) or without (+AHT) tethered TetR-Clr4* Data are mean ±SD (n=3), P<0.05 (t-test).

(B) TetR-Clr4* was completely removed from F0 epe1Δ 4xtetO-ade6+ tetR-Clr4* cells by crossing to epe1Δ lacking TetR-Clr4* and 4xtetO-ade6+. F1 progeny were crossed to epe1Δ cells, generating epe1Δ F2 progeny. epe1+ F2xwt progeny were produced by crossing epe1+ into epe1Δ 4xtetO-ade6+ F2 cells. Naïve epe1Δ 4xtetO-ade6+ cells never expressed TetR-Clr4*. Colony colour, qRT-PCR and qChIP assays to assess silencing and transcription of 4xtetO-ade6+, and H3K9me2 levels on 4xtetO-ade6+ in indicated cell types. Data are mean ±SD (n=3). 4xtetO-ade6+ RNA levels are significantly reduced in F0, F1 and F2 compared to wild-type cells without TetR-Clr4*; P<0.05 (t-test).