Journal: Cancer Science

Article Title: Synthetic disulfide-bridged cyclic peptides mimic the anti-angiogenic actions of chondromodulin-I

doi: 10.1111/j.1349-7006.2012.02276.x

Figure Lengend Snippet: Effects of the synthetic and tailed chondromodulin-I (ChM-I) cyclic peptide on tumor angiogenesis and growth in an animal model in which OUMS-27 cells (5 × 10 6 cells) were subcutaneously inoculated in the backs of 4 week-old nude mice. (A) Time-course of tumor volume changes. When the tumor volume reached about 45 mm 3 , each mouse was injected around the tumor mass each day for the initial 5 days with PBS (50 μL) alone (♢), PBS containing 4.3 nmol (20 μg) ChM-I cyclic peptide with a tail (○), or PBS containing 0.2 nmol (5 μg) recombinant human ChM-I (rhChM-I) (●) (arrows). The tumor volumes were determined by the width 2 × length × 0.52. Values are the means ± SD from at least six animals per group. (B) Gross appearance of tumors excised on day 21. Bar, 10 mm. (C) Immunohistochemical staining of the CD31-positive vasculature. On day 21, tumor tissues were excised, fixed, and cross-sectioned. The sections were then stained with toluidine blue (left panels), and semi-serial sections were stained with an anti-type II collagen antibody ( green signal in the right panels) and an anti-CD31 antibody ( red signal in the right panels), respectively. Bars, 100 μm. (D) The CD31-positive area was measured as described in the methods section. Values are the means ± SD of five tumors per group. * P < 0.05, ** P < 0.01.

Article Snippet: Anti-CD31 antibody and anti-collagen type II antibody were obtained from BD PharMingen (San Diego, CA, USA) and Rockland (Gilvertsville, PA, USA), respectively.

Techniques: Animal Model, Injection, Recombinant, Immunohistochemical staining, Staining

Journal: Nature

Article Title: Capillary cell-type specialization in the alveolus.

doi: 10.1038/s41586-020-2822-7

Figure Lengend Snippet: Fig. 1 | Two stable, intermingled alveolar capillary cell types. a, Alveolar capillaries in adult mouse lung immunostained for PECAM1. b, t-distributed stochastic neighbour embedding (t-SNE) plot of endothelial cell populations annotated in scRNA-seq data for adult mouse lung13. c, Heat map of expression of capillary subset markers and the general endothelial marker Cldn5 in individual capillary cells. CPM, counts per million. d–f, Single-molecule fluorescent in situ hybridization (smFISH) for the capillary subset markers Apln (d, f) or Ednrb and Car4 (aCap) (e), and Aplnr (gCap) (d–f), in adult mouse lung. Images in d (right) and f show individual aCap and gCap cells. g, Relative abundance of aCap cells, gCap cells and cells that co-express aCap and gCap markers (intermediate (IM) cells) in lungs from 3-month-old (young) and 24-month-old (aged) mice (data shown as mean; n = 500 cells scored per mouse; 2 mice per group). h, i, Co-expression of tdTomato lineage label (asterisks) and aCap marker Ednrb but not gCap marker Aplnr (h), or gCap marker Aplnr but not aCap marker Apln (i), in lungs collected six months after mature aCap (h) or gCap (i) cells were lineage-labelled. Blue, DAPI. Scale bars, 10 μm.

Article Snippet: CD34 (BD Biosciences, 347660):https://www.bdbiosciences.com/eu/applications/research/clinical-research/oncology-research/ 3 nature research | reporting sum m ary O ctober 2018 blood-cell-disorders/surface-markers/human/purified-mouse-anti-human-cd34-my10/p/347660 Endomucin (Invitrogen, eBioV.7C7, 14-5851-82): https://www.thermofisher.com/antibody/product/Endomucin-Antibody-cloneeBioV-7C7-V-7C7-Monoclonal/14-5851-82 Integrin alpha8 (R&D, AF4076): https://www.rndsystems.com/products/mouse-rat-integrin-alpha8-antibody_af4076 Pecam1 (rat anti-mouse; BD Biosciences, 553370): https://www.bdbiosciences.com/ds/pm/tds/553370.pdf Pecam1 (mouse anti-human; R&D, BBA7): https://www.rndsystems.com/products/human-cd31-pecam-1-antibody-9g11_bba7 tdTomato (Rockland, 600-401-379): https://rockland-inc.com/store/Antibodies-to-GFP-and-Antibodies-to-RFP-600-401-379O4L_24299.aspx VE-Cadherin (R&D, AF938): https://www.rndsystems.com/products/human-ve-cadherin-antibody_af938 The following primary antibodies were used to stain turtle and/or alligator tissue.

Techniques: Expressing, Marker, In Situ Hybridization

Journal: International Journal of Molecular Sciences

Article Title: Enteropathogenic Escherichia coli (EPEC) Recruitment of PAR Polarity Protein Atypical PKCζ to Pedestals and Cell–Cell Contacts Precedes Disruption of Tight Junctions in Intestinal Epithelial Cells

doi: 10.3390/ijms21020527

Figure Lengend Snippet: Temporal redistribution of TJ proteins and barrier dysfunction caused by EPEC. ( A–C ) SKCO-15 cells were plated on Transwells and infected or not (UI) with EPEC. Total JAM-A, JAM-A S285, JAM-A Y280, occludin, and ZO-1 localization and TER were determined. ( A ) EPEC does not alter the distribution of total JAM-A. In contrast, JAM-A S285 is displaced from the cell–cell contacts to the cytoplasm at 30 min post-infection. Tyrosine phosphorylation of JAM-A Y280 is apparent at 60–120 min post-infection. Scale bars, 10 µm. ( B ) EPEC induces the endocytosis of occludin and ZO-1 at 1 and 2 h post-infection, respectively. Scale bars, 10 µm. ( C ) TER drops significantly as early as 45 min post-infection and progressively decreases over time as more TJ proteins are displaced. TER reported as percent change from baseline. * p < 0.01, *** p < 0.001.

Article Snippet: Par3 (07-330, EMD Millipore), Par6 (ab49776 and ab6022, Abcam, Cambridge, MA, USA), aPKCζ (sc-17781, Santa Cruz Biotechnology, Dallas, TX, USA), p-aPKCζ–T560 (ab62372, Abcam, Cambridge, MA, USA), p-aPKCζ–T410 (sc-12894R, Santa Cruz Biotechnology, Dallas, TX, USA), actin (A2066, Sigma-Aldrich, St. Louis, MO, USA), F-actin BODIPY 558/568 Phalloidin (B3475, Invitrogen, Life Technologies Carlsbad, CA, USA), occludin (33-1500, Invitrogen, Life Technologies, Carlsbad, CA, USA), JAM-A S285 (sc-17430, Santa Cruz Biotechnology, Dallas, TX, USA), JAM-A Y280 (600-401-GN5, Rockland Immunochemicals Inc, Limerick, PA), JAM-A (361700, Invitrogen, Life Technologies, Carlsbad, CA, USA), and ZO-1 (61-7300, Invitrogen, Life Technologies, Carlsbad, CA, USA).

Techniques: Infection, Phospho-proteomics