Journal: Heart and Vessels

Article Title: Cluster analysis of extracellular matrix biomarkers predicts the development of impaired systolic function within 1 year of acute myocardial infarction

doi: 10.1007/s00380-022-02118-8

Figure Lengend Snippet: Exploratory factor analysis results in cohort of AMI patients a The rotated factor matrix of log-transformed ECM biomarkers in 120 AMI patients using EFA. The large ovals represent each latent factor with the percentage of variance for each factor described in bold. The small ovals represent the variables included within each latent factor, and the loading factors are displayed in bold below. b Mann–Whitney U testing demonstrated a significant increase in Factor 1 scores in patients with impaired systolic function compared to preserved systolic function. c No differences were observed in Factor 2 scores between systolic function groups. Median and interquartile range are plotted, and graphs were created using GraphPad Prism software, version 7.04 for Windows. Abbreviations: MMP matrix metalloproteinase, TIMP tissue inhibitor of matrix metalloproteinase, PINP N-terminal type I procollagen

Article Snippet: Levels of PINP (Human PINP ELISA, MyBioSource, California, USA) and TIMP-1 (Human Duoset ELISA, R&D Systems, Minnesota, USA) were measured using ELISA according to manufacturer instructions.

Techniques: Transformation Assay, MANN-WHITNEY, Software

Journal: eLife

Article Title: Piezo1/2 mediate mechanotransduction essential for bone formation through concerted activation of NFAT-YAP1-ß-catenin

doi: 10.7554/eLife.52779

Figure Lengend Snippet: ( a ) Representative images of alizarin red and alcian blue stained limbs from 3 weeks old mice with the indicated genotypes. Bone fractures are indicated with arrows and quantification (n = 4) is shown on the right side. ( b ) Expression of Pieozo 1 and 2. Western blotting analysis of the humerus bone devoid marrow from the Piezo1-Tdtomato mice of the indicated ages. Piezo1 protein levels were shown by RFP immunoblotting. Piezo2 protein was too low to be detected. RT-qPCR analysis of Piezo2 expression of cortical bones from E18.5 and 3 weeks old mice was performed. n = 5–6. ( c ) Masson’s Trichrome staining of humerus sections from P5 mouse pups. Reduction in collagen content (blue staining) was observed in the Piezo1 CKO or Piezo1/2 DKO mutants. Boxed regions are shown in higher magnification in the right panel. ( d ) Detection of serum PINP levels in the 4 weeks old mice with the indicated genotypes by ELISA. Bone formation was most significantly reduced in the Piezo1/2 DKO mutant. (n = 4–7, means ± SD). ( e, f ) Immunostaining of Ctsk ( e ) and Trap1 ( f ) in humerus sections from P0 mouse pups. Boxed regions were shown in higher magnification in the right panel. Osteoclast differentiation was increased in both Piezo1 CKO and Piezo1/2 DKO mutants. ( g ) qPCR analysis of osteoblast and osteoclast gene expression from bone tissues of P0 pups with indicated genotypes (n = 3, means ± SD). *p<0.05, **p<0.01, ***p<0.001, two-tailed unpaired Student’s t -test or one-way ANOVA followed by Tukey’s multiple comparisons tests when ANOVA was significant ( a, d and g ) . Figure 2—figure supplement 1—source data 1. Original numbers used for quantification and Western blots.

Article Snippet: Serum PINP levels were measured using a commercial kit from Immunodiagnostic Systems, according to the manufacturer’s instructions.

Techniques: Staining, Expressing, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Mutagenesis, Immunostaining, Gene Expression, Two Tailed Test

Journal: eLife

Article Title: Piezo1/2 mediate mechanotransduction essential for bone formation through concerted activation of NFAT-YAP1-ß-catenin

doi: 10.7554/eLife.52779

Figure Lengend Snippet: ( a ) Whole mount alizarin red and alcian blue staining of P21 mice with indicated genotypes. Dorsal view of the ribcages was shown in the lower panel. Arrows indicate bone fractures in the ribs. ( b ) Representative 2D μCT images of the femurs from P21 mice with indicated genotypes. The quantified parameters are shown in . ( c ) Expression of Sp7-Cre::GFP in humerus sections of P0 pups with indicated genotypes. ( d ) tdTomato+ cells were sorted from long bones of the Prrx1-Cre;tdTomato fl/+ and Sp7-Cre;tdTomato fl/+ mice and cultured under osteogenic differentiation under static or fluid shear stress (FSS) condition. Piezo1 and Piezo2 expression were analyzed using RT-qPCR. ( e ) Serum PINP levels in 6 weeks old mice of indicated genotypes detected by ELISA. As all bones were affected by the Sp7-GFP::Cre driver, PINP levels were significantly reduced in both Piezo1 or Piezo1/2 mutants (n = 3–4, means ± SD). ( f ) RNA samples from the P0 humerus bone tissues of indicated Prrx1-Cre -driven mutants were subject to RNA seq. Heat-map analysis of differentially expressed genes with fold change increase >2.82 fold and reduction to <0.35 fold. Piezo1 CKO or Piezo1/2 DKO mutants showed similar alteration of gene expression genome wide. ( g ) KEGG pathway analysis of differentially expressed genes. Prominent reductions in Hippo and Wnt signaling were identified (boxed). *p<0.05, **p<0.01, ***p<0.001, one-way ANOVA followed by Tukey’s multiple comparisons tests when ANOVA was significant . Figure 4—source data 1. Differentially expressed genes and orginal numbers for quantification.

Article Snippet: Serum PINP levels were measured using a commercial kit from Immunodiagnostic Systems, according to the manufacturer’s instructions.

Techniques: Staining, Expressing, Cell Culture, Shear, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, RNA Sequencing, Gene Expression, Genome Wide