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. Author manuscript; available in PMC: 2015 Dec 1.
Published in final edited form as: Nat Med. 2015 May 18;21(6):628–637. doi: 10.1038/nm.3866

Figure 1. SMC lineage tracing provides evidence for large populations of phenotypically modulated SMCs within BCAs of 18 Week WD fed SMC eYFP+/+ Apoe−/− mice.

Figure 1

Immunofluorescence staining of representative BCAs from 18 week Western diet fed SMC eYFP+/+ Apoe−/− mice. Results show that differentiated SMCs present at the time of tamoxifen injection (YFP+) subsequently give rise to multiple phenotypes including: (a, b) phenotypically modulated SMCs (YFP+ACTA2− cells highlighted in white rectangle), (c) Mϕ-like SMCs (YFP+ACTA2−LGALS3+), (d) mesenchymal stem cell-like SMCs (YFP+ACTA2−SCA1+), and (e) MF-like cells (YFP+ACTA2+PDGFβR+). Scale bar represents 50μm (a) and 10μm (b-e). (c-e) Yellow arrows indicate de-differentiated (YFP+/ACTA2−) SMCs, white arrows indicate differentiated (YFP+/ACTA2+) SMCs. Samples were either fixed and embedded in paraffin (a-c, e) or Neg40 (d). (f) Flow cytometry of single cell suspension from 18 Week WD fed SMC eYFP+/+ Apoe−/− mouse aortas to further assess phenotypically modulated SMC populations. The gating strategy included: forward versus side scatter gate; a singlets gate; and YFP+ cell gate. Sub-populations of YFP+ cells were found to be double positive for ITGAM (CD11b) and F4/80, ITGAM (CD11b) and ITGAX (CD11c), LGALS3 (mac2), and double positive for SCA1 and ENG after going through lin− gating, n = 6 animals.