Fig 1: IRAR regulates chemokine expression.A Co -expression network of IRAR and differentially expressed mRNAs. The solid line indicates positive regulation and the dotted line indicates negative regulation. B qRT-PCR analysis of IRAR in mTECs transfected with IRAR overexpressing lentivirus (LncRNA-OE) or Negative lentivirus (LncRNA-NC). C–E qRT-PCR analysis of CXCL1 (C), CXCL2 (D), CCL2 (E) in mTECs transfected with IRAR overexpressing lentivirus or Negative lentivirus. F Western blot analysis of CXCL1, CXCL2 and CCL2 in mTECs transfected with IRAR overexpressing lentivirus or Negative lentivirus.. Data are means from three independent experiments. G–J qRT-PCR analysis of IRAR (G), CXCL1 (H), CXCL2 (I) and CCL2 (J) in mTECs. mTECs were transfected with GapmeR IRAR (GapmeR-Hypoxia) or Negative control (Control-Hypoxia) for 48 h, then treated with hypoxia. K RNA fluorescence in situ hybridization (FISH) assays indicated co-expression between IRAR and CXCL1, CXCL2, CCL2 in the kidney at 24 h after ischemia reperfusion. Scale bar, 100 µm. L RNA immunoprecipitation (RIP) experiments were performed using antibodies against CXCL1, CXCL2 and CCL2. RIP enrichment was determined relative to the input controls. M RNA pulldown assays were performed in tubular epithelia cells to examine the association of IRAR with CXCL1, CXCL2 and CCL2. Data represent mean ± SEM. n = 4. **p < 0.01.
Fig 2: Inhibition of IRAR reduces ischemia-reperfusion (IR)-induced production of chemokines and infiltration of immune cells.A–C ELISA of CXCL1 (A), CXCL2 (B), CCL2 (C) in the kidney 24 h after renal IR (n = 6–8). D Immunofluorescence staining of Gr-1 and F4/80 in the kidney 6 h after renal IR. Gr-1 was used as a marker of neutrophils and F4/80 was used as a marker of macrophages. Scale bar, 100 µm (n = 6–8). E Quantification of Gr-1-positive neutrophils and F4/80-positive macrophages in the kidneys. Data represent mean ± SEM. n = 4. *p < 0.05, **p < 0.01.
Fig 3: CXCL2 in omental adipocytes is critical for GC growth in a xenograft model.Control group: AGS cells treated with control media were subcutaneously injected with control media (n = 13). siNT OmAd-CM group: AGS cells treated with siNT OmAd-CM were subcutaneously injected with siNT OmAd-CM (n = 18). siCXCL2 OmAd-CM group: AGS cells treated with siCXCL2 OmAd-CM were subcutaneously injected with siCXCL2 OmAd-CM (n = 5). a Body weight curves. b Representative macroscopic images of resected tumours treated with control media, siNT OmAd-CM and siCXCL2 OmAd-CM in SCID mice. c Tumour growth curve. Tumour size and volume were measured twice per week. d VEGFA expression in tumour tissues. Each graph bar represents the relative ratios of 2-??Ct of siNT OmAd-CM and siCXCL2 OmAd-CM to that of control media. Mean, 1.0 (control), 5.4 (siNT OmAd-CM), 0.1 (siCXCL2 OmAd-CM). e Representative images of Ki67 immunohistochemistry (×200). f Ki67 index. Each bar represents the average rate of Ki67-positive cells in GC tumour tissues (×400). Mean, 4.4 (control), 40.2 (siNT OmAd-CM), 6.5 (siCXCL2 OmAd-CM). g Representative images of tumour angiogenesis (×200). h Quantification of tumour angiogenesis. Each bar represents the average number of CD31-positive microvessels in GC tumour tissues (×400). Mean, 3.3 (control), 29.3 (siNT OmAd-CM), 10.5 (siCXCL2 OmAd-CM).
Fig 4: Ischemia-reperfusion (IR) induces production of chemokines and infiltration of immune cells.A Hierarchical clustering analysis of the differentially expressed mRNAs in IR group compared to Sham group (green, low; red, high). Three significantly upregulated mRNAs: CXCL1, CXCL2, and CCL2 (fold-change > 10) were highlighted in red box. B KEGG pathway enrichment analysis for upregulated mRNAs. C–E Time course of chemokine CXCL1 (C), CXCL2 (D), and CCL2 (E) expression. Data represent mean ± SEM. n = 6–8. **p < 0.01 versus Sham group. F Immunofluorescence staining of Gr-1 and F4/80 in the kidney 6 h and 24 h after renal IR. Gr-1 was used as a marker of neutrophils, and F4/80 was used as a marker of macrophages. Scale bar, 100 µm. G Quantification of Gr-1-positive neutrophils and F4/80-positive macrophages in the kidneys. Data represent mean ± SEM. n = 6. **p < 0.01.
Fig 5: Proposed model of interaction between omental adipocytes and GC growth.CXCL2 secreted from omental adipocytes activates AKT phosphorylation of gastric cancer cells, which directly promotes gastric cancer growth and motility. In addition, VEGFA in gastric cancer cells is also upregulated through HIF1a upregulation, resulting in angiogenesis. Consequently, gastric cancer cells were transformed to a more aggressive phenotype, which induces peritoneal metastasis thorough recruitment to the omentum itself.
Supplier Page from Abcam for Mouse MIP2 ELISA Kit (CXCL2)