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. Author manuscript; available in PMC: 2016 Mar 9.
Published in final edited form as: Cell Rep. 2016 Jan 28;14(5):1018–1024. doi: 10.1016/j.celrep.2016.01.006

Figure 4. Condensin Regulates the Position Effect Within otr Repeats via RITS and is Essential for CENP-A Centromeric Localization.

Figure 4

(A) cut3-477 mutant cells with ura4+ in otr repeats 2, 8 or 11 were serially diluted and spotted on the –ura medium, and incubated at 32°C.

(B) ChIP analysis of H3K9 methylation in otr repeats 2, 8 and 11 in cut3-477. Error bars indicate SD. (*) P < 0.05. n.s., not significant.

(C) Overexpression of Cut3 perturbs the position-dependent silencing in otr repeats. Serial dilution of wild--type cells carrying ura4+ in indicated otr repeat and nmt1-cut3+ were incubated in –ura medium lacking thiamine. OE, overexpression.

(D) ChIP analysis of Chp1-GFP in otr3L repeats 2, 8 or 11 in cut3-477. Error bars indicate SD. (*) P < 0.05. n.s., not significant.

(E) Cut3-myc is co-immunoprecipitated with Chp1-GFP. Extracts from cells carrying Cut3-myc and Chp1-GFP was immunoprecipitated with a GFP antibody. Input and immunoprecipitate were analyzed by western blotting using the antibody against the myc tag.

(F) CENP-Acnp1 is mislocalized in cut3-477 mutant. cut3-477 cells expressing CENP-Acnp1-GFP at endogenous level were incubated at 34°C for 5 hrs. Scale bar, 2 μm.