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. 2014 Jan 30;10(1):e1004115. doi: 10.1371/journal.pgen.1004115

Figure 3. Loss of CHH DNA methylation at the promoter is not sufficient to activate the element but enhances the response to heat stress.

Figure 3

(A) Total CHH methylation at LTRs of ONSEN 1 and 2 (contributing the majority of exDNA) and ONSEN 8 (not activated) in non-stressed and 30 h HS wild type plants. Only CHH positions shared between all three LTRs were considered. CHH methylation profiles along the LTRs and data for individual genomic copies are shown in Supplementary Fig. 2A–F and Supplementary Dataset S1. (B) Total CHH methylation of the same ONSEN LTRs in non-stressed and 30 h HS drm1/drm2/cmt3 triple mutant plants (ddc). Only CHH positions shared between all three LTRs were considered. CHH methylation profiles along the LTRs and data for individual genomic copies are shown in Supplementary Fig. 2G–L and Supplementary Dataset S1. (C) Relative amount of ONSEN RNA determined by quantitative RT-PCR in three week old wild type and ddc mutant plants, non-stressed and after 30 h HS. Bars represent the ONSEN transcripts in relation to that of AtSAND and normalized to WT at 0 h HS. Error bars correspond to the s.e.m. (n = 3). (D) Quantification of ONSEN extrachromosomal DNA from the same samples as in F. Bars represent the ratio between extrachromosomal and integrated copies determined by densitometry after Southern blot analysis. Error bars correspond to the s.d. (n = 3).