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. Author manuscript; available in PMC: 2013 Aug 1.
Published in final edited form as: Nat Methods. 2013 Jan 13;10(2):171–177. doi: 10.1038/nmeth.2332

Figure 4. H3K4dime on the MYH11 promoter persists during phenotypic switching in vivo in SMC lineage tracing mice developing atherosclerosis.

Figure 4

(a) ChIP assay performed in cultured SMCs treated with vehicle (DMSO) or POVPC (10 μg/mL, 24h) showing similar H3K4dime enrichment on the MYH11. Mean s.d.; n = 3. (b) mRNA quantification of MYH11 showing POVPC-induced decrease in MYH11 mRNA level compared with vehicle. Mean ± s.d; n = 3; *P <0.01. (c)Identification of modulated SMCs in SMC-eYFP+/+ ApoE−/− mice fed with Western diet for 18 weeks. Brachiocephalic arteries (BCA) sections of these mice were stained with eYFP (cyan) and MYH11 (red). eYFP+ SMCs were identified within the media where they co-express MYH11 and within the atherosclerotic lesion where they lose expression of MYH11. Scale bar = 100 μm. (d) BCA sections of SMC-eYFP+/+ ApoE−/− mice were stained with eYFP (cyan) and ACTA2 (yellow) and were analyzed for MYH11 H3K4dime PLA.Scale bar = 100 μm.(e). Image corresponding to region 1 boxed in (d) Medial SMCs are ACTA2+ eYFP+ and Myh11 H3K4dime PLA+ (arrows). Higher magnification with eYFP (i), PLA (ii) and merged image with Dapi (iii). (f)Image corresponding to region 2 boxed in (d). Within the atherosclerotic lesion, a substantial fraction of eYFP+ACTA2− cells is Myh11 H3K4dime PLA+ indicating cells of SMC origin not identifiable based on detection of endogenous SMC markers (arrows). Scale bar (e–f) = 10 μm. Higher magnification with eYFP (i), PLA (ii) and merged image with Dapi (iii).