cd144 mouse mab  (Santa Cruz Biotechnology)

Journal: bioRxiv

Article Title: DOT1L-AF10–mediated H3K79me3 promotes NF-κB p65–dependent inflammatory activation in endothelial cells

doi: 10.64898/2026.03.20.713137

Figure Lengend Snippet: (A-B) Immunofluorescence staining of histological sections from partially ligated (D-Flow) and contralateral unligated (S-Flow control) carotid arteries of HFD-fed C57BL/6 mice (n = 3), performed four weeks post-surgery, showing endothelial expression of DOT1L (A) and H3K79me3 (B). EC are marked by CD144 (green), nuclei by DAPI (blue), and target signals (DOT1L or H3K79me3) in red. Fluorescence intensities (AU) in individual EC (dots) from three biological replicates are indicated together with the mean. A total of ≥ 45 EC were analyzed per condition. (C-E) En face preparations were double stained with anti–VE-cadherin (green, EC marker) and an anti-DOT1L antibody (red, C) or H3K79me3 antibody (red, D, E) in both lesser (disturbed flow [D-flow]) and greater (steady laminar flow [S-flow]) curvature areas including TNF-α treated aortic tissue (E). Images were obtained from the luminal surface of the aorta. The levels of nuclear EZH2 and H3K27me3 were analyzed using Image J software. (n = 3) Magnification: x10, x63; Scale: 200 µm, 20 µm. Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by Welch’s t-test.

Article Snippet: For tissue sections, permeabilization was performed using 0.5% Triton X-100, followed by blocking with 10% goat serum supplemented with anti-mouse IgG (1:1000) for 1 h. Cells and tissue sections were incubated overnight at 4°C with the following primary antibodies: DOT1L Rabbit mAb (1:500; #90878), H3K79me3 Rabbit mAb (1:500; #74073), eNOS Rabbit mAb (1:100; #32027), ICAM1 Rabbit pAb (1:500; #4915), NF-κB p65 Rabbit mAb (1:800; #8242, Cell Signaling Technology, Danvers, USA), H3K79me3 Rabbit pAb (1:50; #PA5-96121), AF10 Rabbit pAb (1:500; #BS-3696R), AF17 Rabbit pAb (1:1000; #A302-198A, Invitrogen), and CD144 Mouse mAb (1:25; #SC-9989, Santa Cruz Biotechnology, Dallas, USA).

Techniques: Immunofluorescence, Staining, Control, Expressing, Fluorescence, Marker, Software

Journal: Communications biology

Article Title: Angular difference in human coronary artery governs endothelial cell structure and function.

doi: 10.1038/s42003-022-04014-3

Figure Lengend Snippet: Fig. 6 Increasing bifurcation angles of coronary artery incrementally impacts the EC morphology. FE-SEM (top) based structural and surface level analysis of EC exposed to D-Flow using 30°, 60° and 80° microchannels. Magnification: ×5000, Scale: 10 µm. Flow exposed (4 h; 30°, 60° and 80° microchannels) EA.hy926 cells stained for cell-cell contact markers CD31 (second) and CD144 (penultimate). DAPI staining is shown in blue. Circularity of such cells was assessed using ImageJ. Cell circularity AU values per individual cells are indicated together with mean. Total number of cells, n ≥30. Values represent the mean ± SD. *p < 0.05, **p < 0.01 and ***p < 0.001, by one-way ANOVA. Magnification: ×50/×63, Scale: 80 µm/20 µm. Cell cytoskeletal F-actin (bottom) staining for EA.hy926 exposed to D-Flow (D-variable or D-fixed regions) in 30°, 60° and 80° microchannels, using phalloidin. DAPI staining is shown in blue. Magnification: ×63, Scale: 20 µm.

Article Snippet: Following a 10 min fixation with 4% paraformaldehyde (PFA) and permeabilization using 0.1% Triton X-100 (5 min), the glass slides were blocked with 2% bovine serum albumin for 1 h; and further incubated overnight at 4 °C with eNOS Rabbit mAb (1:100, #32027), ICAM1 Rabbit pAb (1:500; #4915), CD31 (PECAM-1) Mouse mAb (1:1000; #3528) (Cell Signaling Technology, Danvers, USA), and CD144 (VE-Cadherin) Mouse mAb (1:500; #SC-9989; Santa Cruz Biotechnology, Dallas, USA) primary antibodies.

Techniques: Staining