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. Author manuscript; available in PMC: 2021 May 3.
Published in final edited form as: Circ Res. 2021 Feb 24;128(9):1258–1275. doi: 10.1161/CIRCRESAHA.120.318688

Figure 6. Genetic ablation of CARMN affects the area, the volume and the composition of plaques developed in the aortic arches of CARMN-/- versus CARMN+/+ animals.

Figure 6

Schematic representation of CARMN locus located on mouse chromosome 18 and the CRISPR/Cas9 deletion of the first exon and 4.8 kilobases of the promoter (total 5 kilobases). The arrows next to each transcript name, indicate the direction of transcription. The grey arrows and box indicate where common primers to both isoforms were designed. B) Validation of knockout strategy by qRT-PCR using common primer set performed on the aortic arches of CARMN+/+ (n=4) and CARMN-/- (n=6) mice. Mann-Whitney was used to assess statistical significance indicated with p values. C) Schematic workflow used for the induction of mouse atherosclerosis using AAV8-mediated overexpression of PCSK9 and high cholesterol diet in CARMN+/+ and CARMN-/- animals. D) Representative tomographic reconstruction of pictures obtained by Optical Projection Tomography (black picture) and non-tomographic projection of the mouse aortic arch (white picture). The arrows indicate the brachiocephalic artery. Quantification of the plaque volumes in CARMN+/+ (n=7) and CARMN-/- (n=8) animals performed using CTan software. Student’s t-test was used to assess statistical significance indicated with p values. E) Representative Haematoxylin & Eosin staining of cross-sections from brachiocephalic artery of CARMN+/+ and CARMN-/- animals. Pictures acquired at 10X magnification. Scale bar 200μm. F) Quantification of the plaque areas at the site of maximum lesion of brachiocephalic arteries in CARMN+/+ (n=10) and CARMN-/- (n=9) mice, performed using Fiji software. Student’s t-test was used to assess statistical significance indicated with p values. G) Haematoxylin & Eosin staining of cross-sections from aortic roots of wild type and CARMN-/-animals. Pictures acquired at 5X magnification. Scale bar 200μm. H) Quantification of plaque area of aortic roots of CARMN+/+ (n=14) and CARMN-/- (n=15) animals, performed using QuPath software. Student’s t-test was used to assess statistical significance indicated with p values.