Journal: bioRxiv

Article Title: Molecular mechanisms of coronary artery disease risk at the PDGFD locus

doi: 10.1101/2023.01.26.525789

Figure Lengend Snippet: (A) Schematic of experimental design showing that dissected aortic tissues were harvested for single cell RNA sequencing (scRNAseq) and histology analyses from SMC-specific lineage tracing control (Ctl) and lineage tracing Pdgfd knockout (KO) mice. Eight-week-old mice, 2 Ctl and 3 KO captures (two mice per capture), were treated with tamoxifen twice at 3-day intervals and subsequently fed high fat diet for 16 weeks and then sacrificed. Tissues were digested to single cells, tdTomato positive (tdT+) fluorescence and negative (tdT−) cells collected and captured on the10x Chromium controller, libraries generated and sequenced. (B) Uniform manifold approximation and projection (UMAP) of scRNAseq results identified 13 different clusters at 2.6 clustering resolution, with respective biological cluster identities as defined by cluster marker genes. (C) UMAP displaying expression of indicated markers reflecting unique cluster identity: Cnn1 , SMC; Fn1 , FMC; Ibsp , CMC; Rgs5, pericytes. (D) UMAP visualizing dimension reduction plots of Pdgfd and Pdgfrb expression. (E) UMAP images comparing feature expression of tdTomato positive cells from Ctl and KO mice. The dotted line is generated based on the Ctl image. (F) Bar plot presenting the average percentage of lineage traced cells and (G) non-lineage traced cells in Ctl and KO groups.

Article Snippet: Sections were incubated overnight at 4 °C with an anti-Cnn1 rabbit monoclonal primary antibody (1:400 dilution; TA327614; Origene), or a CD68 rabbit polyclonal antibody (1:300 dilution; ab125212; Abcam), after development with Dab, samples were mounted with EcoMount medium (Biocare Medical #EM897L).

Techniques: RNA Sequencing Assay, Control, Knock-Out, Fluorescence, Generated, Marker, Expressing

Journal: bioRxiv

Article Title: Molecular mechanisms of coronary artery disease risk at the PDGFD locus

doi: 10.1101/2023.01.26.525789

Figure Lengend Snippet: (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. (C) Quantification of lesion, and (D) acellular areas in Ctl and KO groups expressed as a ratio of the total vessel area per section. (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. (G) Representative sections stained for Cnn1, a marker of the differentiated SMC. (H) Quantification of Cnn1 positive (Cnn1+) area at the media, and (I) compared to total cross-sectional area expressed as a ratio of the total vessel area per section. (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. (L) Representative images of Col2a1 RNAscope of the aortic root in Ctl and KO mice. (M) Quantitative RNAscope of Col2a1 and (N) Ibsp expression. (O) Representative images stained for calcium deposits with alizarin Red S. (P) Quantification of calcium deposits. Each dot represents quantification from identical level sections from individual animals. Data expressed as mean ± s.e.m with p -values using an unpaired t -test. ** p <0.01, *** p <0.001.

Article Snippet: Sections were incubated overnight at 4 °C with an anti-Cnn1 rabbit monoclonal primary antibody (1:400 dilution; TA327614; Origene), or a CD68 rabbit polyclonal antibody (1:300 dilution; ab125212; Abcam), after development with Dab, samples were mounted with EcoMount medium (Biocare Medical #EM897L).

Techniques: Staining, Activity Assay, Expressing, Marker, Immunohistochemistry, RNAscope

Journal: Animals : an Open Access Journal from MDPI

Article Title: Prostacyclin Synthesis and Prostacyclin Receptor Expression in the Porcine Myometrium: Prostacyclin Potential to Regulate Fatty Acid Transporters, Cytokines and Contractility-Related Factors

doi: 10.3390/ani12172237

Figure Lengend Snippet: Full names of genes exploited to examine the mRNA transcript abundance with real-time PCR.

Article Snippet: CNN1 , Calponin 1 , Ss03392449_g1.

Techniques:

Journal: Animals : an Open Access Journal from MDPI

Article Title: Prostacyclin Synthesis and Prostacyclin Receptor Expression in the Porcine Myometrium: Prostacyclin Potential to Regulate Fatty Acid Transporters, Cytokines and Contractility-Related Factors

doi: 10.3390/ani12172237

Figure Lengend Snippet: Effect of prostaglandin I2 (PGI2; prostacyclin) analog, iloprost, on the mRNA expression of contractility-related genes (panel A ; GJA1, OXTR, CNN1, and CALD1 ) and genes encoding transporters (panel B ) of glucose ( SCL5A1 ), amino acids ( SLC38A1 ), and fatty acids ( CD36 and SLC27A4 ) in the myometrium. Myometrial explants were incubated for 8 h without (control) or with 1 µM of iloprost. Values from real-time PCR for each studied mRNA transcript were normalized to geometric averaging of GAPDH and HPRT1 mRNA expression. Data are expressed as the mean ± SEM ( n = 6). Asterisk specifies the difference compared with the control value (* p < 0.05).

Article Snippet: CNN1 , Calponin 1 , Ss03392449_g1.

Techniques: Expressing, Incubation, Control, Real-time Polymerase Chain Reaction

Journal: Experimental and Therapeutic Medicine

Article Title: Calcimimetic R568 reduced the blood pressure and improved aortic remodeling in spontaneously hypertensive rats by inhibiting local renin-angiotensin system activity

doi: 10.3892/etm.2018.6734

Figure Lengend Snippet: Immunohistochemical analysis of proliferating remodeling protein expression in the thoracic aorta of rats. Representative images of (A) Calponin; (B) α-SMA, smooth muscle α-actin; (C) OPN, osteopontin; (D) PCNA, proliferating cell nuclear antigen. Quantification of (E) Calponin; (F) α-SMA, smooth muscle α-actin; (G) OPN, osteopontin; (H) PCNA, proliferating cell nuclear antigen. Data are means ± standard deviation (n=7). SHR+NS groups vs. the age-matched WKY+NS groups, *P<0.05; SHR16w+NS groups vs. SHR8w+NS groups, # P<0.05; SHR+R568 groups vs. the age-matched WKY+R568 groups, **P<0.05; SHR16w+R568 groups vs. SHR8w+R568 groups, ## P<0.05; SHR+R568 groups vs. SHR+NS groups, a P<0.05. WKY, Wistar Kyoto rats; SHR, spontaneously hypertensive rats; NS, normal saline; R568, NPSR568; α-SMA, smooth muscle α-actin; OPN, osteopontin; PCNA, proliferating cell nuclear antigen. Scale bar, 50 µM.

Article Snippet: The sections were incubated with primary mouse polyclonal antibodies against CaSR (1:50; Abcam, MA, USA), calponin (1:600; Boster, Wuhan, China), smooth muscle actin α (α-SMA, 1:400; Boster), proliferating cell nuclear antigen (PCNA, 1:200; Boster), and primary rabbit monoclonal antibody against osteopontin (OPN, 1:100; Abcam) overnight at 4°C.

Techniques: Immunohistochemical staining, Expressing, Standard Deviation, Saline

Journal: Experimental and Therapeutic Medicine

Article Title: Calcimimetic R568 reduced the blood pressure and improved aortic remodeling in spontaneously hypertensive rats by inhibiting local renin-angiotensin system activity

doi: 10.3892/etm.2018.6734

Figure Lengend Snippet: Western blot analysis of proliferating remodeling protein expression in the thoracic aorta of rats. (A) Western blot analysis of the protein expression. (B) Calponin. (C) α-SMA, smooth muscle α-actin. (D) OPN, osteopontin. (E) PCNA, proliferating cell nuclear antigen. Data are means ± standard deviation (n = 7). SHR+NS groups vs. the age-matched WKY+NS groups, *P<0.05; SHR16w+NS groups vs. SHR8w+NS groups, # P<0.05; SHR+R568 groups vs. the age-matched WKY+R568 groups, **P<0.05; SHR16w+R568 groups vs. SHR8w+R568 groups, ## P<0.05; SHR+R568 groups vs. SHR+NS groups, a P<0.05. WKY, Wistar Kyoto rats; SHR, spontaneously hypertensive rats; NS, normal saline; R568, NPSR568; α-SMA, smooth muscle α-actin; OPN, osteopontin; PCNA, proliferating cell nuclear antigen.

Article Snippet: The sections were incubated with primary mouse polyclonal antibodies against CaSR (1:50; Abcam, MA, USA), calponin (1:600; Boster, Wuhan, China), smooth muscle actin α (α-SMA, 1:400; Boster), proliferating cell nuclear antigen (PCNA, 1:200; Boster), and primary rabbit monoclonal antibody against osteopontin (OPN, 1:100; Abcam) overnight at 4°C.

Techniques: Western Blot, Expressing, Standard Deviation, Saline