Journal: Cellular and molecular biology (Noisy-le-Grand, France)

Article Title: Homocysteine modulates CXCL10/CXCR3 axis activity to induce endothelial dysfunction.

doi: 10.14715/cmb/2024.70.2.28

Figure Lengend Snippet: Fig. 3. (A, B) qRT-PCR and Western blot analysis of CXCL10 and CXCR3 expression in HAECs treated with different concentrations of Hcy. (C) IHC analysis of the aorta in HHcy mice showing CXCL10 and CXCR3 expression. (D) qRT-PCR analysis of CXCL10 and CXCR3 expression in arterial endothelial cells.

Article Snippet: Additionally, HAECs were exposed to specific agents including Anti-CXCL10 antibodies (701225, Invitrogen, USA), Anti-CXCR3 antibodies (ab71864, Abcam, UK), IgG control antibodies (31154, Thermo Fisher, USA), NBI-74330 (a CXCR3 inhibitor, 4528, Tocris Bioscience, UK), and CXCL10 agonist (266-IP, R&D Systems, USA).

Techniques: Quantitative RT-PCR, Western Blot, Expressing

Journal: bioRxiv

Article Title: Neutrophils promote CXCR3-dependent itch in the development of atopic dermatitis

doi: 10.1101/653873

Figure Lengend Snippet: A. Log 2 fold change (Day 8 MC903 vs. EtOH) of Th2 genes in skin from uninjected wild-type, aGR1-treated, and TSLPR -/- animals. B. Log 2 fold change (Day 8 MC903 vs. EtOH) of chemokine genes in skin from uninjected wild-type, aGr1-treated, and TSLPR -/- animals. C. Log 2 fold change (Day 8 MC903 vs. EtOH) of activity-induced genes in trigeminal ganglia from uninjected wild-type, aGr1-treated, and TSLPR -/- animals. D. Log 2 fold change (Day 8 MC903 vs. EtOH) of Lcn2 and activity-induced genes in spinal cord from uninjected and aGr1-treated wild-type mice on day 8. For Figure 4A-D , exact values and corrected p -values are reported in Supplemental Data. E. Quantification of innervation (see Methods) of MC903 and EtOH-treated mouse skin as determined from BTIII staining ( p = 0.8985; two-tailed t-test ( t =0.1294; df =18); n = 9,11 images each from 2 mice per treatment. Exact values are reported in Figure 4-source data 1 . F. CXCL10 levels in skin homogenate as measured by ELISA on day 8 of the MC903 model for uninjected animals (left; * p = 0.029 ( t =2.715, df =7); two-tailed t-test; n = 4,5 animals), animals which received aGr1 for 8 days (middle; p = 0.43 ( t =0.815, df =11); two-tailed t-test; n = 6,6 animals), and TSLPR -/- animals (right; * p = 0.0357 ( t =2.696, df =6); two-tailed t-test; n = 4,4 animals. Skin homogenates were isolated on separate days and so uninjected, WT samples were not compared to aGr1-treated samples or to TSLPR -/- samples. G. (Left) Time spent scratching over a thirty minute interval on days 5, 8, and 12 of the MC903 model, one hour after mice were injected with either 3.31 mM of the CXCR3 antagonist AMG 487 or vehicle (20% HPCD in PBS; 50 µL s.c. in rostral back); (two-way ANOVA: **** p treatment < 0.0001, F(1,67) = 50.64; Tukey’s multiple comparisons: * p day 5 = 0.0216, n=8,10 mice; *** p day 8 = 0.0007, n=18,21 mice; **** p day 12 < 0.0001, n=8,8 mice). (Right) Time spent scratching over a thirty minute interval one hour after mice were injected with either 3.31 mM of the CXCR3 antagonist AMG 487 or vehicle (20% HPCD in PBS; 50 µL s.c. in rostral back), and immediately after mice were injected with 50 mM chloroquine (20 µL i.d., cheek). p = 0.92 ( t =0.0964, df =8); two-tailed t-test; n = 5,5 mice. Values from bar plots in Figures 4F-G are displayed in Figure 4-source data 2 .

Article Snippet: CXCL10 protein was quantified using the Mouse CXCL10 Duoset ELISA kit (R&D Systems; #DY466-05) according to manufacturer’s instructions.

Techniques: Activity Assay, Staining, Two Tailed Test, Enzyme-linked Immunosorbent Assay, Isolation, Injection

Journal: bioRxiv

Article Title: Neutrophils promote CXCR3-dependent itch in the development of atopic dermatitis

doi: 10.1101/653873

Figure Lengend Snippet: A. AD induction first results in increased protease expression and barrier dysfunction, which drives production of the cytokines TSLP and CXCL1 via PAR2 activation within keratinocytes. CXCL1 can recruit neutrophils via its receptor CXCR2. Neutrophils may evoke itch by multiple pathways, including degranulation and release of proteases and histamine, production of sensitizing lipids such as PGE 2 and LTB 4 , 52 and induction of CXCL10 expression, which can activate sensory neurons via CXCR3. TSLP activates a number of immune cells to elicit IL-4 production, including basophils, which results in increased IL-4, recruitment of CD4 + T cells, and sensitization of neurons to promote itch later in the model.

Article Snippet: CXCL10 protein was quantified using the Mouse CXCL10 Duoset ELISA kit (R&D Systems; #DY466-05) according to manufacturer’s instructions.

Techniques: Expressing, Activation Assay