Fig 1: The spinal localization of CXCR2 and its ligand CXCL3 in naive and CCI-exposed rats. Immunofluorescent staining was performed on paraffin-embedded 7 µm (A,B) co-staining of CXCR2 (green) and neuronal marker NeuN (red; upper row); astroglial marker GFAP (red, middle row), and microglial marker IBA1 (red, bottom row). (C,D) co-staining of CXCL3 (green) and neuronal marker NeuN (red; upper row); astroglial marker GFAP (red, middle row), and microglial marker IBA1 (red, bottom row). White arrows indicate representative CXCL3-positive cells that co-localize with IBA1-positive cells. Scale bar for all pictures: 25 µm.
Fig 2: Effects of the repeated administration of NVP CXCR2 20 (NVP; 10 µg/5 µl; i.t.; 16 h and 1 h before CCI and then once a day for 7 days) on the protein levels of CXCR2, IBA1, GFAP, CXCL1, CXCL2, and CXCL3 proteins (A–I) in the spinal cord (A–F) and DRG (G–I) on the 7th day after CCI in rats. The data are presented as the mean fold changes relative to the control ± SEM (5–6 samples per group). Intergroup differences were analyzed using ANOVA with Bonferroni's multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001 indicate differences vs. naive rats. #p < 0.05, indicate differences between V-treated and NVP-treated rats. CCI, chronic constriction injury; N, naive; V, vehicle; NVP, NVP CXCR2 20.
Fig 3: Effects of single administrations of CXCL1, CXCL2, and CXCL3 (A–F) on nociceptive transmission in naive mice. The effects of single intrathecal administrations of CXCL1, CXCL2, CXCL3 (2, 400, or 800 ng/5 µl) on mechanical hypersensitivity (von Frey test, A–C) and thermal hypersensitivity (cold plate test, D–F) were measured at 1.5, 5, and 24 h after administration. Data are presented as the means ± SEM (6 mice per group). The results were evaluated using one-way ANOVA, followed by Bonferroni's test for comparisons of selected pairs measured separately at each time point. *p < 0.05, **p < 0.01, ***p < 0.001 for the comparison of vehicle-treated naive animals with all groups at the indicated time points. Additionally, the results were evaluated using two-way ANOVA to determine the time × drug interaction (please see results in Chapter 3.3). V, vehicle.
Fig 4: CXCL1, CXCL2, CXCL3, CXCR1, and IL-1ß expression in carcinoma cells xenografted into nude mice at 1, 2, and 3 weeks, respectively.
Fig 5: Western blot analysis of tumor and peritumor tissues from the CBRH-7919 cell line demonstrated the up-regulation of CXCL1, CXCL2, CXCL3, and XCR1 and down-regulation of CXCR1 in tumor tissues at 1, 2, and 3 weeks. T: Tumor tissue, A: Peritumor tissue.
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