Journal: Communications Biology

Article Title: Porcine intraepithelial lymphocytes undergo migration and produce an antiviral response following intestinal virus infection

doi: 10.1038/s42003-022-03205-2

Figure Lengend Snippet: a Sankey diagram depicts the chemokine secretion in the host cells and the intestinal mucosa infected with variant and classical PEDV strains. b The protein expression of CCL2 in the ligated terminal jejunum injected with PEDV and in the jejunum from piglets orally inoculated with PEDV was detected via immunofluorescence analysis. CCL2 and PEDV were immunolabeled with anti-CCL2 mAb (green) and anti-PEDV polyclonal antibody (Red), respectively; the cell nuclei were stained with DAPI (blue). Scale bars, 50 μm. c The protein expression of CCL2 and CCL5 in the medium of Marc-145 cells was detected using ELISA kits. d Schematic of experimental setting used to study the migration of IELs influenced by chemokines induced by PEDV infection. e Chemokines CCL2, CCL5, and the CCL2 inhibitor TC1 were added into the medium of the basolateral side. Filters from the co-culture system were determined via CLSM, and the intraepithelial and transepithelial (DAPI, blue) migration of IELs (CFSE, green) in response to different treatments was shown by a three-dimensional (3D) rendering of representative fields. f Quantitative analysis of transepithelial IELs was performed. The number of transepithelial IELs was counted from five random fields of view at a unit area (0.078 mm 2 ) for each of the three individual filters. g The IELs/IPEC-J2 co-culture model was pretreated with an CCL2 inhibitor TC1 for 2 h (DMSO as a negative control) by upper compartment inoculation, followed by a culture with PEDV-infected or whole inactivated PEDV-treated Marc-145 cells. The intraepithelial and transepithelial (DAPI, blue) migration of IELs (CFSE, green) was detected via CLSM. h Quantitative analysis of transepithelial IELs in each experimental group. All data are the mean ± SD, and comparisons were performed using one-way ANOVA. * P < 0.05, ** P < 0.01. The results are from at least three different experiments.

Article Snippet: To inhibit the CCR2 receptor activity of IELs, we used CCR2 antagonist 4 (Teijin compound 1, TC1; MedChemExpress, China).

Techniques: Infection, Variant Assay, Expressing, Injection, Immunofluorescence, Immunolabeling, Staining, Enzyme-linked Immunosorbent Assay, Migration, Co-Culture Assay, Negative Control

Journal: Oncotarget

Article Title: Targeting CCR2 with its antagonist suppresses viability, motility and invasion by downregulating MMP-9 expression in non-small cell lung cancer cells

doi: 10.18632/oncotarget.16837

Figure Lengend Snippet: (A) Culture medium from LC99A, LC11-18, NCI-H460 and A549 cell lines was collected and CCL2 level was measured by ELISA; (B) mRNA expression of CCR2 in NCI-H460 and A549 cells were determined by RT-PCR; (C) the protein level of CCR2 in NCI-H460 and A549 cell were detected by Western blot.

Article Snippet: Besides the limitation on the cost, CCL2 antibodies have shown some limitation for cancer treatment, as the removal of anti-CCL2 antibody in cancer treatment was found leading to an increased metastatic burden in an orthotropic mammary tumor model [ ], While the cessation of CCR2 antagonist (RDC018, GlaxoSmithKline) did not enhance malignant progression in hepatocellular carcinoma postsurgical recurrence mice models.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Oncotarget

Article Title: Targeting CCR2 with its antagonist suppresses viability, motility and invasion by downregulating MMP-9 expression in non-small cell lung cancer cells

doi: 10.18632/oncotarget.16837

Figure Lengend Snippet: (A) A549 cells were treated with 0-200 ng/ml CCL2 for 72h and then subjected to cell viability assay; (B) A549 cells were treated with 50 ng/ml CCL2 for 72h with or without pretreatment of CCR2 antagonist (CAS 445479-97-0) (0-20 nM). Cell viability was then measured by WST-1 assay. Bars, SD ( *, p<0.05, **, p<0.01) .

Article Snippet: Besides the limitation on the cost, CCL2 antibodies have shown some limitation for cancer treatment, as the removal of anti-CCL2 antibody in cancer treatment was found leading to an increased metastatic burden in an orthotropic mammary tumor model [ ], While the cessation of CCR2 antagonist (RDC018, GlaxoSmithKline) did not enhance malignant progression in hepatocellular carcinoma postsurgical recurrence mice models.

Techniques: Viability Assay, WST-1 Assay

Journal: Oncotarget

Article Title: Targeting CCR2 with its antagonist suppresses viability, motility and invasion by downregulating MMP-9 expression in non-small cell lung cancer cells

doi: 10.18632/oncotarget.16837

Figure Lengend Snippet: (A) Transwell migration assay. A549 cells were set in upper chamber of a 24-well transwell. RPMI 1640 with 10% FCS culture medium containing a serial dilution of CCL2 was added to the bottom well. After 4 h incubation, cells were fixed and the migration cell numbers for each group were counted. (B) Wound healing scratch assay. A549 cells were pretreated with or without CCR2 antagonist, CAS 445479-97-0 (10 nM) for 2 h. Then an artificial scratch wound was created using a p10 pipet tip. Cells were then incubated with serum free medium containing variable concentration of CCL2 and monitored at indicated time points. (C) Representative photographs of transwell migration assay with A549 cells in the presence of CCL2 (100 ng/ml) with or without pretreatment of CAS 445479-97-0. (D) The migration cell numbers for each group were calculated and are shown. The difference (* P < 0.05) was analyzed by Two-tailed paired Student's t-test.

Article Snippet: Besides the limitation on the cost, CCL2 antibodies have shown some limitation for cancer treatment, as the removal of anti-CCL2 antibody in cancer treatment was found leading to an increased metastatic burden in an orthotropic mammary tumor model [ ], While the cessation of CCR2 antagonist (RDC018, GlaxoSmithKline) did not enhance malignant progression in hepatocellular carcinoma postsurgical recurrence mice models.

Techniques: Transwell Migration Assay, Serial Dilution, Incubation, Migration, Wound Healing Assay, Concentration Assay, Two Tailed Test

Journal: Oncotarget

Article Title: Targeting CCR2 with its antagonist suppresses viability, motility and invasion by downregulating MMP-9 expression in non-small cell lung cancer cells

doi: 10.18632/oncotarget.16837

Figure Lengend Snippet: A549 cells were pretreating with 10 nM CCR2 antagonist (CAS 445479-97-0) for 24 h or 5uM MMP-9 inhibitor (sc-311437) for 30 min, followed by stimulation with CCL2 for 24 h. The protein level of MMP-9 was then detected by Western blot (A) , the migration and invasion activity of A549 cells were assessed by transwell (B) and Matrigel assays (C) , respectively. * p < 0.05 represents statistically significant differences between the group pretreated with CCR2 antagonist or MMP-9 inhibitor and the CCL2-treated group.

Article Snippet: Besides the limitation on the cost, CCL2 antibodies have shown some limitation for cancer treatment, as the removal of anti-CCL2 antibody in cancer treatment was found leading to an increased metastatic burden in an orthotropic mammary tumor model [ ], While the cessation of CCR2 antagonist (RDC018, GlaxoSmithKline) did not enhance malignant progression in hepatocellular carcinoma postsurgical recurrence mice models.

Techniques: Western Blot, Migration, Activity Assay