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. 2010 Jun 10;6(6):e1000986. doi: 10.1371/journal.pgen.1000986

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Vrbsky et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) The chart shows the frequency and distribution of cytosine methylation in the 1L-0' region. In total, 30 clones from five independently treated wild-type samples and 19 clones from three independent rdr2 samples were analyzed. Asterisks indicate the third cytosine in the CCCTAAA sequence. (B) The proportion of methylated cytosines in the 1L-0' region of wild-type plants depending on the position within the telomeric repeat as determined by bisulfite sequencing. (C) Frequency of cytosine methylation in the whole 1L-0' region according to sequence context in wild-type and rdr2 plants. (D) Cytosine methylation in bulk telomeric DNA assayed by dot-blot hybridization. Bisulfite-treated (BS) genomic DNA was spotted onto a membrane (∼7, 33 and 200 ng from each sample) and sequentially hybridized with AAAATTT, TTTAGGG and CCCTAAA probes. Untreated wild-type genomic DNA, and untreated and bisulfite-treated (BS) plasmids carrying ∼750 nt of Arabidopsis telomeric DNA were used as controls. (E,F) Quantification of signals obtained with oligo (E) and long (F) TTTAGGG probes. The signal intensity of non-converted DNA (obtained with the TTTAGGG probe) was normalized to the amount of telomeric DNA determined from hybridization with the CCCTAAA probe. The signal from BS-treated plasmid served to determine background hybridization of the probes to fully converted non-methylated telomeric DNA. Error bars represent standard deviation (N = 3).