Journal: Frontiers in Cell and Developmental Biology

Article Title: NRP2 as an Emerging Angiogenic Player; Promoting Endothelial Cell Adhesion and Migration by Regulating Recycling of α5 Integrin

doi: 10.3389/fcell.2020.00395

Figure Lengend Snippet: NRP2 is not regulated by ITGB3 during VEGFR2-mediated signaling or during migration over FN. (A) : WT and β3HET ECs were plated onto FN and incubated for 48 h at 37°C and 5% CO 2 . EC extracts were immunoblotted using antibodies to NRP2, ITGB3 and HSC70. N = 3 independent experiments, **** P < 0.0001. (B,C) : siRNA-transfected ECs were plated onto FN and incubated for 48 h at 37°C and 5% CO 2 . ECs were subsequently starved in serum-free media and stimulated with VEGF (30 ng/ml) for the indicated timepoints. EC extracts were immunoblotted using antibodies to phospho-VEGFR2, VEGFR2, NRP2, NRP1, HSC70, phospho-ERK and ERK. N = 6 independent experiments, P > 0.05 (nsd). Left panel shows representative Western blot images, right panel shows densitometric analysis of band intensities normalized against HSC70 and obtained using ImageJ TM . Asterisks indicate statistical significance, nsd indicates no significant difference from unpaired two-tailed t -tests. (C) : siRNA-transfected ECs were seeded onto FN and incubated for 3 h at 37°C and 5% CO 2 . Fixed images were taken every 10 min for 15 h at 37°C and 5% CO 2 using an inverted Zeiss Axiovert microscope with one-phase contrast. Random migration speed was quantified using the ImageJ TM plugin mTrackJ in μm/hour. N = 4 independent experiments, n ≥ 30 ECs per experimental condition, **** P < 0.0001.

Article Snippet: Primary antibodies (all used at 1:1000 dilution and purchased from Cell Signaling Technology, unless noted otherwise) were: anti-NRP2 (clone D39A5), anti-ITGB3 (clone 4702S), anti-HSC70 (clone B-6, Santa Cruz Biotechnology), anti-phospho VEGFR2 (Y1175) (clone 2478), anti-VEGFR2 (clone 2479), anti-NRP1 (clone 3725S), anti-phospho ERK1/2 (clone 9101), anti-ERK1/2 (clone 4695), anti-ITGA5 (clone 4705S), anti-ITGB1 (clone ab179471, Abcam), anti-clathrin heavy chain-1 (clone ab21679, Abcam), anti-caveolin-1 (clone ab18199, Abcam), anti-Rab11 (clone 3539), ERG: Ab92513, Pecam-1 (77699), prox-1 (clone ab11941, Abcam), claudin-5 (clone ab131259, Abcam), VE-cadherin (clone ab205336, Abcam).

Techniques: Migration, Incubation, Transfection, Western Blot, Two Tailed Test, Microscopy

Journal: Bioactive Materials

Article Title: Cellulose fibers-reinforced self-expanding porous composite with multiple hemostatic efficacy and shape adaptability for uncontrollable massive hemorrhage treatment

doi: 10.1016/j.bioactmat.2020.12.014

Figure Lengend Snippet: a . SEM images of CMCP after absorbing protein. Scale bars: 2 μm. b . Measurement of the number of platelets adhered on different materials. c . SEM images of the adhered and activated platelets on the composite. d . The absorbance of the hemoglobin-containing solution. e . Blood cells adhesion (platelets and RBCs), platelets activation, and fibrin network formed and trapped blood cells. Yellow dotted circles indicate the RBCs, red dotted rectangles indicate the activated platelets. *p < 0.05 and # p < 0.05 represent significant difference compared with the blank group. f , g , h . The fluorescence intensity and relative content of Ca2 + in platelets, CD61 and CD62p were measured to evaluate the stimulating effects of CMCP on platelets.

Article Snippet: 50 μL of FITC (Shanghai Macklin Biochemical Co., Ltd) labeled anti-CD61 and anti-CD62p (Sino Biological, China) were added after dilution, and the mixture was kept in the dark at 4 °C for 20 min.

Techniques: Activation Assay, Fluorescence

Journal: NPJ Precision Oncology

Article Title: Targeted single-cell proteomic analysis identifies new liquid biopsy biomarkers associated with multiple myeloma

doi: 10.1038/s41698-023-00446-0

Figure Lengend Snippet: IMC multiplexed proteomic analysis panel of metal-labeled antibodies.

Article Snippet: CD61 , Mouse_IgG1_VI-PL2 , Bi209 , Fluidigm , 981514 , 3209001B.

Techniques: