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. 2021 Apr 29;118(3):772–784. doi: 10.1093/cvr/cvab156

Figure 3.

Figure 3

SMC Drebrin inhibits atherosclerosis and SMC-to-foam cell transdifferentiation. Common carotid arteries from Dbnflox/flox and SMC-Dbn−/− mice were transplanted into the right common carotid of congenic Apoe−/− mice as interposition grafts, and harvested 4 weeks later. (A) Sections were stained with a modified connective tissue stain to facilitate planimetry of neointima and media. Scale bars = 100 μm. (B) Neointimal, medial, and total arterial cross-sectional areas were measured by planimetry (ImageJ), and plotted for nine distinct carotid grafts/genotype along with means ± SE. Compared with Dbnflox/flox: *P < 0.01 (Mann–Whitney). (C) Serial sections of carotid grafts from A were incubated with anti-CD68 or isotype control IgG, along with Hoechst 33342 (DNA). Negative control IgG yielded no colour (not shown). Scale bars = 50 μm. L, lumen; IEL, internal elastic lamina. (D) In cross sections from C, the CD68-positive neointimal area was divided by the cognate total neointimal area, and plotted for nine distinct carotid grafts/genotype, along with means ± SE. Compared with Dbnflox/flox: *P < 10−3 (t test). (E) Serial sections of carotid grafts were incubated simultaneously with BODIPY® 493/503 and goat anti-apoE, followed by Hoechst 33342 (DNA) and anti-goat/Alexa 546 IgG. Confocal microscopy used an optical slice thickness of 1 μm. Serial sections stained with non-immune primary IgG yielded no colour (not shown). The dotted white lines indicate the IEL. The dashed boxes indicate areas enlarged further in the adjacent panels. L, lumen. Scale bars = 20 μm. Co-localization of red (apoE) with either blue (Hoechst) or green (BODIPY) was performed as in Figure 2. (F) BODIPY-stained material was judged to be cellular as in Figure 2. Foam cell prevalence was determined as in Figure 2 and plotted for 9 distinct carotids/genotype, along with means ± SE. Compared with Dbnflox/flox: *P < 10−3 (Mann–Whitney). (G) BODIPY+ neointimal or medial cells (≥100 per layer per carotid graft) were scored as containing yellow (apoE+/BODIPY+) or not; the percentage of apoE+/BODIPY+ in each layer was plotted for nine distinct carotids per genotype, along with means ± SE. Compared with Dbnflox/flox: *P < 10−3 (t test).