Journal: Oncology Letters

Article Title: Study on the mechanism of oral administration of tetrandrine during neoadjuvant chemotherapy for colon cancer

doi: 10.3892/ol.2023.13811

Figure Lengend Snippet: Expression levels of CCL5, CXCL2 and CXCL10 in colon cancer tissue culture supernatant. Expression levels of chemotactic factors, (A) CCL5, (B) CXCL2 and (C) CXCL10 in surgically resected fresh colon cancer tissue culture supernatant were detected with ELISA. *P<0.05, **P<0.01 compared with the control group. CCL, chemokine ligand; CXCL, chemokine (C-X-C) motif ligand; Control group, patients not receiving tetrandrine; tet group, patients receiving tetrandrine.

Article Snippet: ELISA was performed with the following kits, according to the manufacturer's instructions: Human TNF-α ELISA kit (cat. no. ab181421; Abcam), human IL-1β ELISA kit (cat. no. ab100562; Abcam), human IL-6 ELISA kit (cat. no. ab46027; Abcam), human IL-15 ELISA kit (cat. no. ab218266; Abcam), human chemokine ligand (CCL)2 ELISA kit (cat. no. ab179886; Abcam), human CCL20 ELISA kit (cat. no. ab178015; Abcam), human CCL5 ELISA kit (ab174446; Abcam), human chemokine (C-X-C) motif ligand (CXCL) 1 ELISA kit (cat. no. ab190805; Abcam), human CXCL2 ELISA kit (cat. no. ab184862; Abcam), human CXCL3 ELISA kit (cat. no. ab234574; Abcam), human CXCL5 ELISA kit (cat. no. ab212163; Abcam) and human CXCL10 ELISA kit (cat. no. ab83700; Abcam).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: PLoS ONE

Article Title: MMP-9 and CXCL8/IL-8 Are Potential Therapeutic Targets in Epidermolysis Bullosa Simplex

doi: 10.1371/journal.pone.0070123

Figure Lengend Snippet: A. SQRT-PCR shows increased matrix metalloproteinase mRNA expression in EBS-DM cell lines (n = 4). B. MMP-9 ELISA of 48-h-conditioned cell culture supernatant shows 2-fold upregulation of MMP-9 in KEB-7 and 46-fold upregulation in EBDM-1 at the protein level (n = 4). C. MMP-9 levels were highly increased in EBS patients blister fluids compared to healthy controls (n = 3 to n = 4). The numbers correlate with . Student`s t -test was performed with p values: * ≤0.05, ** ≤0.01, *** ≤0.005, ΔΔ≤0.0005, ΔΔΔ≤0.0001. (In 2C, the Student’s t -test compared the entire patient group to the entire control group).

Article Snippet: Protein levels of KLK5, MMP7, MMP9, CXCL1, CXCL8/IL-8, CXCL11 and CXCL14 were determined in 48-h-conditioned cell culture supernatant and in patients blister fluids by using Quantikine® ELISA (Human KLK5, # DKK500, R&D Systems; Human MMP7, # DMP700, R&D Systems; Human MMP9, # DMP900, R&D Systems; Human CXCL1/GROα, # DGR00, R&D Systems; Human CXCL8/IL-8, # D8000C, R&D Systems; Human CXCL11/I-TAC, # DCX110, Human CXCL14/BRAK, # DY866, R&D Systems) following the manufacturer’s protocol.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Control

Journal: PLoS ONE

Article Title: MMP-9 and CXCL8/IL-8 Are Potential Therapeutic Targets in Epidermolysis Bullosa Simplex

doi: 10.1371/journal.pone.0070123

Figure Lengend Snippet: A. SQRT-PCR of chemokine mRNA expression in NEB-1, KEB-7 and EBDM-1 (n = 3 to n = 5). CXCL1 , CXCL8/IL-8 and CXCL14 expression was increased in KEB-7. Only CXCL11 and CXCL14 were increased in EBDM-1. B. CXCL8/IL-8 ELISA of 48-h-conditioned cell culture supernatant showed 2-fold upregulation at the protein level in KEB-7 but not in EBDM-1, which correlates with the SQRT-PCR results (n = 4). C. CXCL8/IL-8 concentrations were highly increased in EBS patients blister fluids. In blister fluids of healthy controls no CXCL8/IL-8 was detectable (n = 3 to n = 4). The numbers correlate with . Student’s t -test was performed with p values: * ≤0.05, *** ≤0.005, Δ≤0.001, ‡ = no significant difference between investigated cell lines. (In 6C, the Student’s t -test compared the entire patient group to the entire control group).

Article Snippet: Protein levels of KLK5, MMP7, MMP9, CXCL1, CXCL8/IL-8, CXCL11 and CXCL14 were determined in 48-h-conditioned cell culture supernatant and in patients blister fluids by using Quantikine® ELISA (Human KLK5, # DKK500, R&D Systems; Human MMP7, # DMP700, R&D Systems; Human MMP9, # DMP900, R&D Systems; Human CXCL1/GROα, # DGR00, R&D Systems; Human CXCL8/IL-8, # D8000C, R&D Systems; Human CXCL11/I-TAC, # DCX110, Human CXCL14/BRAK, # DY866, R&D Systems) following the manufacturer’s protocol.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Control

Journal: Journal of clinical & cellular immunology

Article Title: The Anti-inflammatory Effect of Alpha-1 Antitrypsin in Rhinovirus-infected Human Airway Epithelial Cells

doi: 10.4172/2155-9899.1000475

Figure Lengend Snippet: Increased IL-8 production in cultured COPD brushed airway epithelial cells. Brushed airway epithelial cells from COPD patients (n=6) and normal subjects (n=6) were cultured under air-liquid interface (ALI) condition for 10 days. After 24 hours of air exposure, IL-8 was measured by ELISA. Data are expressed as means ± SEM.

Article Snippet: Protein levels of human IL-8, or mouse KC were determined by using a human IL-8 or a mouse KC DuoSet ELISA kit (R&D Systems, Minneapolis, MN) as per manufacturer’s instruction.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Acetylcholine in the gingival epithelium drives the pathogenesis of periodontitis

doi: 10.3389/fcell.2025.1701252

Figure Lengend Snippet: Acetylcholine orchestrates a multi-dimensional defense program in epithelial cells. (A) Network of GSEA-GO enrichment analysis for Pg vs. NC DEGs. Each node represents a GO term, with circle size and color indicating the pathway size and the NES score, respectively. Node-sharing clusters represent functionally related term groups. (B) Transcript counts of CXCL and CCL family genes in HOKs RNA-seq data. Only genes with non-zero counts across all 12 samples are displayed, with orange bars indicating the mean. (C) Volcano plot of DEGs in Pg vs. NC comparison. Colored dots represent significantly differential expression (|log2 fold change| > 1, adjusted p -value <0.05). (D) Volcano plot of PD vs. HC DEGs in epithelial cells from single-cell data. Colored dots represent significantly differential expression (|log2 fold change| > 1, adjusted p -value <0.05). (E) Enzyme-linked immunosorbent assay (ELISA) results of HOK supernatants for CXCL1 and CXCL8. *, p < 0.05 and **, p < 0.01. (F) Enrichment analysis of DEGs from the Pg + Ach vs. Pg groups using the fgsea function, showing four significantly enriched functional categories [adjusted p < 0.05, normalized enrichment score (NES) > 1.5].

Article Snippet: The levels of CXCL1 and CXCL8 were then measured using an enzyme-linked immunosorbent assay (ELISA) kit (Proteintech, KE00133 and KE00453), according to the manufacturer’s instructions.

Techniques: RNA Sequencing, Comparison, Quantitative Proteomics, Enzyme-linked Immunosorbent Assay, Functional Assay

Journal: British journal of cancer

Article Title: Selective inhibition of HDAC6 promotes bladder cancer radiosensitization and mitigates the radiation-induced CXCL1 signalling.

doi: 10.1038/s41416-023-02195-0

Figure Lengend Snippet: Fig. 5 Tubacin represses RT-induced oncogenic CXCL1 signalling. a CXCL1 mRNA expression in T24 cells with or without 5 μM tubacin (Tub) treatment followed by 5 Gy irradiation (RT) was assessed by qPCR. Panobinostat (PAN, 10 nM) was used for comparison. b CXCL1 protein levels in conditioned medium from T24 cells with or without 5 μM Tub treatment followed by 5 Gy irradiation were measured by ELISA at 24 h. c Migration and invasion assays of T24 cells with or without 5 μM Tub treatment followed by 5 Gy irradiation. Migrated and invaded cells on the lower Transwell membrane were stained and calculated at 16 and 20 h, respectively. Representative images are shown below individually. d Migration and invasion assays of T24 cells treated with conditioned medium (CM) from cells with or without 5 μM Tub treatment followed by 5 Gy irradiation. Migrated and invaded cells on the lower Transwell membrane were stained and calculated at 16 and 20 h, respectively. Representative images are shown below individually. e Migration and invasion assays of Tub- and RT + Tub-treated T24 cells in the presence of 10 μg/mL anti-CXCL1 or control IgG treatment. Panobinostat (PAN)- and RT + PAN-treated cells were used for comparison. Migrated and invaded cells on the lower Transwell membrane were stained and calculated at 16 and 20 h, respectively. Representative images are shown below individually. f Western blotting for H3K9ac, acetyl-α-tubulin and Snail in Tub- and RT + Tub-treated T24 cells with or without anti-CXCL1 treatment at 24 h. Panobinostat (PAN)- and RT + PAN-treated cells were used for comparison. Snail protein expression was quantified by ImageJ. Data are presented as the means ± SD from three independent biological replicates. Student’s t-test was used for significant differences. *P < 0.05; **P < 0.01; ***P < 0.001. g Immunohistochemical staining for CXCL1 in tumour tissues obtained from urothelial carcinoma patients. Representative images are shown, and brown precipitates indicate positive signals. Scale bar represents 50 μm. h Overall survival analysis of 40 urothelial carcinoma patients categorised by CXCL1 protein levels. Statistical differences were assessed by the log-rank (Mantel–Cox) test. i Categorisation of CXCL1 expression in patients with high (pT3-4) and low (pT1-2) T stages.

Article Snippet: The conditioned medium was collected at 24 h and the CXCL1 protein level was determined using a Quantikine ELISA kit (Human CXCL1/GROα, R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions.

Techniques: Expressing, Irradiation, Comparison, Enzyme-linked Immunosorbent Assay, Migration, Membrane, Staining, Control, Western Blot, Immunohistochemical staining

Journal: Nature Communications

Article Title: Snail promotes ovarian cancer progression by recruiting myeloid-derived suppressor cells via CXCR2 ligand upregulation

doi: 10.1038/s41467-018-03966-7

Figure Lengend Snippet: CXCR2 ligands are highly expressed in Snail-high ovarian tumors. a GO (Gene Ontology) term analysis of DNA microarray data obtained from HM-1-control ( n = 3) and HM-1-shSnail (sh1; n = 2 and sh2; n = 2) cells. Genes that were significantly downregulated were used for GO term analysis. Genes are listed in Supplementary Table . b Snail expression was correlated with the expression of CXCR2 ligands (CXCL1; upper left, P < 0.0001, CXCL2; upper right, P < 0.001, and CXCL5; lower left, P < 0.05) in TCGA samples ( n = 266). Pearson’s product-moment correlation analysis. c Reverse transcription polymerase chain reaction (RT-PCR) of human ovarian cancer cell lines, OVCAR8 and OVCAR8-shSnail (left), and OVCA433 and OVCA433-Snail (right); n = 4. d , e ELISA of cell supernatants of human ovarian cancer cell lines, d OVCAR8 and OVCAR8-shSnail, and e OVCA433 and OVCA433-Snail; n = 6. f RT-PCR of HM-1-control and HM-1-shSnail cells; n = 5. g , h Levels of CXCL1, CXCL2, and CXCL5 in subcutaneous tumors ( g ) and in blood ( h ) taken from mice bearing HM-1-control and HM-1-shSnail tumors; n = 6. * P < 0.05, ** P < 0.01, and *** P < 0.001 (unpaired t -test in c – e , g , h ; one-way ANOVA with Tukey’s multiple comparisons test in f ). Averaged data are presented as the mean ± SEM

Article Snippet: Mouse CXCL1, CXCL2, and CXCL5 protein levels were measured by mouse Quantikine ELISA kits for CXCL1, CXCL2, or CXCL5 (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s protocols.

Techniques: Microarray, Control, Expressing, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Nature Communications

Article Title: Snail promotes ovarian cancer progression by recruiting myeloid-derived suppressor cells via CXCR2 ligand upregulation

doi: 10.1038/s41467-018-03966-7

Figure Lengend Snippet: Snail induces CXCL1 and CXCL2 via the NF-κB pathway and possibly via the direct binding to their promoters. a Western blotting of nuclear Snail, phosphorylated p65 (phospho-p65), p65 and RelB from HM-1, OVCAR8 and OVCA433 cells. HDAC1 was used as control. b Reverse transcription polymerase chain reaction (RT-PCR) to analyze the expression of Cxcl1 and Cxcl2 in HM-1-control and HM-1-shSnail cells (left), and the expression of CXCL1 and CXCL2 in OVCAR8-control and OVCAR8-shSnail cells (right), treated with or without BAY11-7082 (NF-κB inhibitor) at 10 μM for 24 h; n = 4. c Schematic representation of CXCL1 and CXCL2 promoter organization, and the corresponding luciferase reporter constructs pGL-CXCL1 (1523 bp: −1523 to +110 bp, 984 bp: −984 to +110 bp, 300 bp: −300 to +110 bp) and pGL-CXCL2 (1606bp: −1606 to +104 bp, 942 bp: −942 to +104 bp, 457 bp: −457 to +104 bp). TSS transcriptional start site, E1 exon1, and Luc luciferase. The black bars indicate E-boxes (CANNTG), which are the binding sites of Snail. d Luciferase reporter assays to analyze the activity of the pGL-CXCL1 and pGL-CXCL2 promoter constructs in 293FT-control and 293FT-shSnail cells. Relative luciferase activities are shown; n = 5. * P < 0.05, ** P < 0.01, and *** P < 0.001 (one-way ANOVA with Tukey’s multiple comparisons test in b ; unpaired t -test in d ). Averaged data are presented as the mean ± SEM

Article Snippet: Mouse CXCL1, CXCL2, and CXCL5 protein levels were measured by mouse Quantikine ELISA kits for CXCL1, CXCL2, or CXCL5 (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s protocols.

Techniques: Binding Assay, Western Blot, Control, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Expressing, Luciferase, Construct, Activity Assay

Journal: Nature Communications

Article Title: Snail promotes ovarian cancer progression by recruiting myeloid-derived suppressor cells via CXCR2 ligand upregulation

doi: 10.1038/s41467-018-03966-7

Figure Lengend Snippet: CXCR2 ligands induce myeloid-derived suppressor cell infiltration. a Chemotaxis of mouse MDSCs, from subcutaneous tumor of HM-1 tumor-bearing mice, in response to CXCL1, CXCL2, and CXCL5; n = 4. b Chemotaxis of mouse MDSCs, from subcutaneous tumor of HM-1 tumor-bearing mice, pre-treated with SB265610 (CXCR2 antagonist) at each concentration in the presence of each CXCR2 ligands (at 100 ng/mL); n = 4. c Chemotaxis response of MDSCs, from human ovarian cancer ascites, to CXCL1, CXCL2, and CXCL5. The chemotaxis index is shown; n = 4. d Chemotaxis response of human MDSCs treated with SB265610 at each concentration in the presence of each CXCR2 ligands; n = 4. * P < 0.05, ** P < 0.01, and *** P < 0.001 (one-way ANOVA with Tukey’s multiple comparisons test in a – d )

Article Snippet: Mouse CXCL1, CXCL2, and CXCL5 protein levels were measured by mouse Quantikine ELISA kits for CXCL1, CXCL2, or CXCL5 (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s protocols.

Techniques: Derivative Assay, Chemotaxis Assay, Concentration Assay

Journal: Nature Communications

Article Title: Snail promotes ovarian cancer progression by recruiting myeloid-derived suppressor cells via CXCR2 ligand upregulation

doi: 10.1038/s41467-018-03966-7

Figure Lengend Snippet: Elevated levels of serum CXCL1 and CXCL2 reflect intratumoral MDSCs and are associated with poor prognosis in ovarian cancer patients. a Serum concentration of CXCL1 (left) and CXCL2 (right) in ovarian cancer patients ( n = 26) vs. that in healthy donors ( n = 8); mean ± SEM; *** P < 0.001 by unpaired t -test. b Correlation between CXCL1 and CXCL2 serum concentrations in ovarian cancer patients ( n = 26); P = 0.0002, R = 0.67. Pearson’s product-moment correlation analysis. c Correlation between serum CXCL1 (left; P = 0.018) and CXCL2 (right; P = 0.09) levels and infiltration of CD33 + cells in peritoneal disseminations; n = 12; Pearson’s product-moment correlation analysis. d Correlation between serum CXCL1 (left; P = 0.049) and CXCL2 (right; P = 0.080) levels and Snail staining scores; n = 12; Pearson’s product-moment correlation analysis. e Overall survival of ovarian cancer patients, comparing the high-serum CXCL1 group ( n = 12) to the low-serum CXCL1 group ( n = 14). Cut-off levels, 42.83 pg/ml; AUC = 0.6842. Hazard ratio, 15.08; 95% confidence interval (CI), 3.859–102.9; P = 0.0005 by log-rank test. f Overall survival of ovarian cancer patients, comparing the high-serum CXCL2 group ( n = 13) to the low-serum CXCL2 group ( n = 13). Cut-off levels, 93.57 pg/ml; AUC = 0.6767. Hazard ratio, 13.87; 95% confidence interval (CI), 3.56–89.72; P = 0.0008 by log-rank test

Article Snippet: Mouse CXCL1, CXCL2, and CXCL5 protein levels were measured by mouse Quantikine ELISA kits for CXCL1, CXCL2, or CXCL5 (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s protocols.

Techniques: Concentration Assay, Staining