αcxcr4  (R&D Systems)

Journal: BMC Complementary and Alternative Medicine

Article Title: Morus alba and active compound oxyresveratrol exert anti-inflammatory activity via inhibition of leukocyte migration involving MEK/ERK signaling

doi: 10.1186/1472-6882-13-45

Figure Lengend Snippet: Effect of oxyresveratrol on CXCR4 signaling. ( A ) Jurkat cells were pre-treated with oxyresveratrol (2.5 μg/ml), resveratrol (2.5 μg/ml) or DMSO vehicle for 1 h at 37°C. After extensive washing, the cells were stained with αCXCR4 (1 μg/ml) and FITC-conjugated secondary antibody (1 μg/ml) or not. The expression level of CXCR4 on the cell membrane is shown in the histogram. Data are representative examples of 3 experiments. ( B & C ) The cells, pre-treated with oxyresveratrol or DMSO, were stimulated with SDF-1 (100 ng/ml) for 0 to 15 min. Cell lysates were analyzed by Western blot with primary antibodies (1 μg/ml) against MAPKs ( B ), MEK1/2 ( C ) or their phosphorylated forms ( B & C ) plus secondary antibodies (0.3 μg/ml). The ratio was obtained by normalizing the signal of the phosphorylated protein to that of the total protein. ( D ) A schematic model describing the mechanism by which oxyresveratrol present in M. alba can inhibit inflammation.

Article Snippet: RPMI 1640 medium, PSQ solution (penicillin, streptomycin and glutamine), sodium pyruvate, non-essential amino acids and HEPES were purchased from Gibco (CA, USA). αCXCR4, FITC-conjugated secondary antibody (Life Technologies, NY, USA) and SDF-1 (R&D systems, MN, USA) were purchased.

Techniques: Staining, Expressing, Western Blot

Journal: Frontiers in Immunology

Article Title: The Activity of the Neutral Sphingomyelinase Is Important in T Cell Recruitment and Directional Migration

doi: 10.3389/fimmu.2017.01007

Figure Lengend Snippet: Neutral sphingomyelinase (NSM) is essential for CXCR4, pERM, and Cdc42 polarization of T cells on FN. Primary control and NSM KD T cells were stimulated with 100 ng/mL SDF-1α, settled onto FN-coated slides for 1 h, stained for Cdc42 (A) , pERM (B) , and CXCR4 (C) , and redistribution toward the leading edge (LE) or uropod, as indicated in the graphs, was analyzed manually. Arrowheads mark CXCR4 or Cdc42 accumulations. Images show representative cells, the graphs (right) show quantification of re-localization with means and SD for n = 3 independent experiments with approximately 30 cells each, average knockdown efficiency: 85.67%. Scale bars, 5 µm.

Article Snippet: For immunostaining, cells were fixed in formaldehyde (4% in PBS), permeabilized (0.1% Triton X-100) and stained for αCdc42 (clone P1), αCXCR4 (clone 12G5) (both Santa Cruz Biotech, Dallas, TX, USA) and αpERM (clone 41 A3, Cell Signaling, Danvers, MA, USA).

Techniques: Control, Staining, Knockdown