Journal: Pediatric research

Article Title: Angiopoietin-1 protects against endotoxin-induced neonatal lung injury and alveolar simplification in mice.

doi: 10.1038/s41390-021-01544-0

Figure Lengend Snippet: Fig. 2 ANGPT2 is upregulated by LPS in HPMEC and directly mediates EC immune. activation, which is attenuated by rhANGPT1. HPMEC were treated with LPS (100 ng/mL), recombinant human ANGPT2 (rhANGPT2, 25 ng/mL), and rhANGPT1 (200 ng/mL), with lysates. used for assays. a ANGPT1, ANGPT2, and TIE2 gene expression by qPCR 24 h after LPS. (n = 4/condition, *p < 0.05, **p < 0.01.) b Quantification of INOS, ICAM1, IL-8, and IL-6 expression by qPCR 6 h after rhANGPT2. (n = 4/condition, **p < 0.01, ***p < 0.001.) c, d HPMEC were treated for 24 h with rhANGPT1 and rhANGPT2, and lysates were used to quantify gene expression of IL-1β, ICAM1, IL-8, and TNF-α (c) and ANGPT2 (d) by qRT-PCR. (n = 3/condition, *p < 0.05, **p < 0.01, ***p < 0.001 between Control and rhANGPT2; rhANGPT2 and rhANGPT1 + rhANGPT2; rhANGPT1 and rhANGPT1 + rhANGPT2.) e ANGPT2 protein was quantified by immunoblot of HPMEC lysates 24 h after rhANGPT1 and rhANGPT2 treatments, with densitometry shown (f). (n = 4/condition, ***p < 0.001 between Control and LPS; LPS and LPS + rhANGPT1). ANOVA (with Tukey) or Mann–Whitney tests were used.

Article Snippet: Recombinant human ANGPT1 (rhANGPT1, 200 ng/mL) and recombinant human ANGPT2 (rhANGPT2, 25 ng/mL) were purchased from R&D Systems.

Techniques: Activation Assay, Recombinant, Gene Expression, Expressing, Quantitative RT-PCR, Control, Western Blot, MANN-WHITNEY

Journal: Journal of cellular biochemistry

Article Title: Mesenchymal Stem Cells Respond to Hypoxia by Increasing Diacylglycerols

doi: 10.1002/jcb.25292

Figure Lengend Snippet: Effect of hypoxia and D609 on secretion of angiogenic factors. After culture of MSC for 48 h in either normoxia or hypoxia and in the presence or absence of 50 μM D609, cells were counted and supernatants were collected and levels of angiogenic factors determined by ELISA. Values represent the amount of angiogenic factor (VEGF, IL-8, or Ang-2) secreted by 1,000 cells over 48 h. Individual symbols represent single experiments performed with MSC derived from a different donor while bars represent the overall averages. Statistically differences where determined by paired Student’s t tests comparing the indicated conditions, with P values indicated over respective brackets.

Article Snippet: For semi-quantitative detection of mRNA levels of VEGF, IL-8, Ang-2, and GAPDH (internal control) real time PCR (RT-PCR) was done using TaqMan primers/probes (Life Technologies): Hs00900055_m1, Hs00174103_m1, Hs01048042_m1, and Hs99999905_m1, respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: Hypoxia-induced mitogenic factor (FIZZ1/RELMα) induces endothelial cell apoptosis and subsequent interleukin-4-dependent pulmonary hypertension

doi: 10.1152/ajplung.00279.2013

Figure Lengend Snippet: HIMF and IL-4 activate VCAM-1 and angiopoietin 2 (Ang2) expression in mouse PMVECs. A–C: primary mouse PMVECs were treated with or without HIMF (20 nM) or rmIL-4 (50 ng/ml) for 24 h. A: VCAM-1 protein expression was analyzed in cell lysates (left). The ratio of VCAM-1 to β-actin is shown (right). ***P < 0.001 vs. vehicle control; n = 3/group. B: cells were stained with fluorescent antibodies to Ang2 and vWF. DAPI was used to stain the nuclei. Images are shown at ×630 magnification. C: Ang2 gene expression was analyzed by quantitative RT-PCR. *P < 0.05 and **P < 0.01 vs. control (n = 3/group). These experiments were repeated at least three times with similar results.

Article Snippet: Probes and primers were designed and synthesized by Applied Biosystems: mu_gapdh (Mm99999915_g1); mu_actb (Mm00607939_s1); mu_pgk1 (Mm00435617_m1); mu_Ccl5 (Mm01302428_m1); mu_Angpt2 (Mm00545822_m1); mu_Pdgfa (Mm01205760_m1); mu_Mmp2 (Mm00439498_m1); mu_Vcam1 (Mm01320970_m1): mu_Timp2 (Mm00441825_m1); and mu_Bmp4 (Mm01332882_m1).

Techniques: Expressing, Staining, Quantitative RT-PCR

Journal: The EMBO Journal

Article Title: Distinct roles of VE ‐cadherin for development and maintenance of specific lymph vessel beds

doi: 10.15252/embj.201798271

Figure Lengend Snippet: RNA isolation and quantitative real‐time PCR (qPCR) using TaqMan Assays

Article Snippet: Gene symbol Reference Angpt1 Mm01129232_m1 Angpt2 Mm00545822_m1 Cdh2 Mm01162497_m1 Cldn5 Mm00727012_s1 Ctgf Mm01192933_g1 Cyr61 Mm00487498_m1 Esam Mm00518378_m1 F11r (Jam1) Mm00554113_m1 Jam2 Mm00470197_m1 Jam3 Mm00499214_m1 Ocln Mm00500912_m1 Pecam1 Mm01242584_m1 Tek Mm00443243_m1 Tjp1 (Zo‐1) Mm00493699_m1 Trp73 Mm00660220_m1 Vegf‐C Mm01202430_m1 Vegf‐D Mm01131929_m1 Wwtr1 (Taz) Mm01289583_m1 Yap1 Mm01143263_m1 Open in a separate window RNA isolation and quantitative real‐time PCR (qPCR) using TaqMan Assays

Techniques: Isolation, Real-time Polymerase Chain Reaction