Fig. 5: In situ studies of mouse atherosclerosis reveal that Pdgfd KO lessens SMC cell state transitions and inflammation but without impact on plaque burden.
From: Molecular mechanisms of coronary artery disease risk at the PDGFD locus
a X-gal staining visualizing β-galactosidase activity (lacZ, blue precipitate) to determine the cellular location of Pdgfd expression in mouse model atherosclerosis. Aortic root sections were also stained with a generic nuclear marker nuclear fast red (NFR), immunohistochemistry for the Cd68 macrophage marker or Cnn1 marker for SMC identification. b Quantification of total vessel area, n = 25 control and 17 KO mouse sections. c Quantification of lesion, n = 26 control and 17 KO mouse sections, and d acellular areas in Ctl and KO groups expressed as a ratio of the total vessel area per section, n = 10 control and 10 KO mouse sections, p < 0.0001. e Representative images identifying expression of the tdTomato gene to visualize the SMC lineage traced cells in aortic sections. f Quantification of tdTomato positive (tdT+) area relative to total vessel area, n = 18 control and 14 KO mouse sections, p < 0.0001. g Representative sections stained for Cnn1, a marker of the differentiated SMC. h Quantification of Cnn1 positive (Cnn1+) area at the media, n = 25 control and 17 KO mouse sections, p = 0.0061 and i compared to total cross-sectional area expressed as a ratio of the total vessel area per section, n = 25 control and 17 KO mouse sections, p = 0.0042. j Representative images of Cd68-stained aortic root area to quantify monocyte recruitment. k Quantification of Cd68 positive (Cd68+) area relative to the vessel area, n = 23 control and 17 KO mouse sections, p = 0.004. l Representative images of Col2a1 RNAscope of the aortic root in Ctl and KO mice. m Quantitative RNAscope of Col2a1, n = 12 control and 12 KO mouse sections, p = 0.0052 and n Ibsp expression, n = 13 control and 13 KO mouse sections, p = 0.0074. o Representative images stained for calcium deposits with alizarin red S. p Quantification of calcium deposits, n = 9 control and 9 KO mouse sections, p = 0.0076. Each dot represents quantification from identical level sections from individual animals. Data expressed as mean ± s.e.m with p-values using a two-sided unpaired t-test.
