Journal: Cells

Article Title: p53 Promotes Cytokine Expression in Melanoma to Regulate Drug Resistance and Migration

doi: 10.3390/cells11030405

Figure Lengend Snippet: Reduced cytokine secretion in p53 knockout melanoma lines. ( A ) Comparative analysis of ligands abundance in supernatant from p53 ko and p53 WT LOX-IMVI cells by multiplex cytokine array. Differential expression of secreted ligand for each knockout clone compared to wild type is shown in the heatmap (as in log2-fold change). The observed ligand concentration (pg/mL) in p53 WT cells are depicted on the right side by numbers >10, 1–10, and <1. Cytokines analyzed by qPCR (in E and E) are indicated by an asterisk on the left side. ( B ) Expression of a subset of the ligands in wild-type and knockout clones by multiplex analysis is demonstrated. n = 3 (mean ± S.D), except for LIF, in which two biological samples had shown the value below the detection limit. ( C ) Validation of CXCL1 expression in p53 ko LOX-IMVI and SK-MEL-5 cells by Elisa analysis; n = 3 (mean ± S.D). Significance was determined by t -test in comparison to the wild-type group, and the observed level of significance is shown by asterisk (* < 0.05; ** < 0.005; *** < 0.0005; **** < 0.0001).

Article Snippet: Human CXCL1 expression was analyzed using Elisa set (R & D system, DY 275), according to company instruction.

Techniques: Knock-Out, Multiplex Assay, Quantitative Proteomics, Concentration Assay, Expressing, Clone Assay, Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Communications biology

Article Title: Single-cell transcriptomes reveal a molecular link between diabetic kidney and retinal lesions.

doi: 10.1038/s42003-023-05300-4

Figure Lengend Snippet: Fig. 4 Transcriptomes of HMCs and HRPCs in response to control or AGEs treatment. a The overlap of highly expressed genes in the normal control groups of HMCs and HRPCs. b Shared enriched GO pathways between HMCs and HRPCs, sorted by p-value. c Volcano plot shows DEGs in HMCs and HRPCs treated with AGEs. The red dots indicate upregulated genes while blue dots indicate downregulated genes. d Expression levels of CXCL1, CXCL2, CXCL3, CXCL5, CXCL6 and CXCL8 were measured by real-time PCR and ELISA between control and AGEs groups. e Significantly enriched KEGG pathways of DEGs in both HMCs and HRPCs treated with AGEs, the size of circle represents genes number enriched in pathways, the darker color of circle indicates a smaller p-value. f The chemokine scores of MCs and RPCs in wt and db/db mice. g The proportions of infiltrating neutrophils and macrophages in the glomeruli of wt and db/db mice. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: Chemokines in the cell culture supernatant were assayed using enzyme-linked immunosorbent assay (ELISA) kits (EK0722, EK0728, EK0359 and EK0413 from BOSTER, Wuhan, China and EK1264 and EK1265 from MULTI SCIENCES, Hangzhou, China) following the manufacturer’s instructions.

Techniques: Control, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Cells

Article Title: Association Between Synovial NTN4 Expression and Pain Scores, and Its Effects on Fibroblasts and Sensory Neurons in End-Stage Knee Osteoarthritis

doi: 10.3390/cells14060395

Figure Lengend Snippet: Primer sequences.

Article Snippet: Furthermore, CXCL1 was determined using an ELISA kit (ProteinTech, Chicago, IL, USA, Cat. No. KE00133), and CXCL6 was measured with another ELISA kit (ProteinTech, Cat. No. KE00274).

Techniques: Sequencing

Journal: Cells

Article Title: Association Between Synovial NTN4 Expression and Pain Scores, and Its Effects on Fibroblasts and Sensory Neurons in End-Stage Knee Osteoarthritis

doi: 10.3390/cells14060395

Figure Lengend Snippet: qPCR analysis of vehicle and recombinant Netrin-4 treated fibroblastic cells derived from the synovium. Relative expression levels of ( A ) NTN4 , ( B ) UNC5B , ( C ) NEO1 , ( D ) MMP1 , ( E ) MMP3 , ( F ) MMP13 , ( G ) VCAM1 , ( H ) CXCL1 , ( I ) CXCL6 , ( J ) IL6 , and ( K ) IL8 following rh-NTN4 treatment compared to vehicle control is presented using box-and-whisker plots. These plots depict the median, quartiles, and range of each dataset. Statistical significances between groups are clearly indicated with lines, and asterisks (*) denote p -values less than 0.05, highlighting statistically significant differences.

Article Snippet: Furthermore, CXCL1 was determined using an ELISA kit (ProteinTech, Chicago, IL, USA, Cat. No. KE00133), and CXCL6 was measured with another ELISA kit (ProteinTech, Cat. No. KE00274).

Techniques: Recombinant, Derivative Assay, Expressing, Control, Whisker Assay

Journal: Cells

Article Title: Association Between Synovial NTN4 Expression and Pain Scores, and Its Effects on Fibroblasts and Sensory Neurons in End-Stage Knee Osteoarthritis

doi: 10.3390/cells14060395

Figure Lengend Snippet: ELISA analysis of cell supernatant in vehicle- and recombinant Netrin-4-treated fibroblastic cells derived from the synovium. Concentrations of ( A ) MMP-1, ( B ) MMP-3, ( C ) MMP-13, ( D ) CXCL1, ( E ) CXCL6, ( F ) IL-6, and ( G ) IL-8 in cell supernatant are presented in box-and-whisker plots, showing the median, 25th and 75th percentiles, and range. * p < 0.05.

Article Snippet: Furthermore, CXCL1 was determined using an ELISA kit (ProteinTech, Chicago, IL, USA, Cat. No. KE00133), and CXCL6 was measured with another ELISA kit (ProteinTech, Cat. No. KE00274).

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Derivative Assay, Whisker Assay

Journal: BioMed Research International

Article Title: The Serum Levels of the Soluble Factors sCD40L and CXCL1 Are Not Indicative of Endometriosis

doi: 10.1155/2016/2857161

Figure Lengend Snippet: Distribution of pain scores measured with the visual analogue scale (VAS), with values from 0 to 10 in endometriosis patients and controls plus serum levels of sCD40L and CXCL1 (numbers represent raw values).

Article Snippet: The CXCL1 serum levels were measured using the commercially available human CXCL1/GRO alpha Quantikine ELISA Kit in concentrations as pg/mL (Cat. number DGR00 Quantikine ELISA Human CXCL1/GRO α Immunoassay, R&D Systems).

Techniques:

Journal: BioMed Research International

Article Title: The Serum Levels of the Soluble Factors sCD40L and CXCL1 Are Not Indicative of Endometriosis

doi: 10.1155/2016/2857161

Figure Lengend Snippet: Patients with endometriosis did not show a statistically significant change in the serum levels of both sCD40L and CXCL1. Boxplots showing the comparison between the levels of expression of sCD40L (a) and CXCL1 (b) in serum of patients with endometriosis versus controls. The levels are presented as log 10 and the p values are indicated above each plot. Arrows next to the boxplots indicate mean ± standard deviation.

Article Snippet: The CXCL1 serum levels were measured using the commercially available human CXCL1/GRO alpha Quantikine ELISA Kit in concentrations as pg/mL (Cat. number DGR00 Quantikine ELISA Human CXCL1/GRO α Immunoassay, R&D Systems).

Techniques: Comparison, Expressing, Standard Deviation

Journal: BioMed Research International

Article Title: The Serum Levels of the Soluble Factors sCD40L and CXCL1 Are Not Indicative of Endometriosis

doi: 10.1155/2016/2857161

Figure Lengend Snippet: The sCD40L and CXCL1 secretion is disease stage independent. Levels of sCD40L (a) and CXCL1 (b) in the control group and based on the different stages of endometriosis classified by rAFS stage in the endometriosis patients, with no statistically significant differences. C: control group, I/II: endometriosis rAFS minimal-to-mild disease, and III/IV: endometriosis rAFS moderate-to-severe disease.

Article Snippet: The CXCL1 serum levels were measured using the commercially available human CXCL1/GRO alpha Quantikine ELISA Kit in concentrations as pg/mL (Cat. number DGR00 Quantikine ELISA Human CXCL1/GRO α Immunoassay, R&D Systems).

Techniques:

Journal: BioMed Research International

Article Title: The Serum Levels of the Soluble Factors sCD40L and CXCL1 Are Not Indicative of Endometriosis

doi: 10.1155/2016/2857161

Figure Lengend Snippet: The serum levels of sCD40L and CXCL1 are not differentially regulated during the menstrual cycle in patients with endometriosis versus controls. Boxplots showing levels of sCD40L (a) and CXCL1 (b) among endometriosis patients and controls by menstrual cycle phases, with no statistically significant differences. CP: control proliferative phase, CS: control secretory phase, EP: endometriosis proliferative phase, and ES: endometriosis secretory phase.

Article Snippet: The CXCL1 serum levels were measured using the commercially available human CXCL1/GRO alpha Quantikine ELISA Kit in concentrations as pg/mL (Cat. number DGR00 Quantikine ELISA Human CXCL1/GRO α Immunoassay, R&D Systems).

Techniques:

Journal: BioMed Research International

Article Title: The Serum Levels of the Soluble Factors sCD40L and CXCL1 Are Not Indicative of Endometriosis

doi: 10.1155/2016/2857161

Figure Lengend Snippet: Patients with deep infiltrating endometriosis (DIE) show a tendency of reduced sCD40L levels and no changes in the levels of CXCL1 in serum when compared to controls. Boxplots showing levels of sCD40L (a) and CXCL1 (b) among controls and endometriosis patients with DIE. The number of samples for each group is given in brackets on the x -axis and the p values of the comparison between the groups are shown below each graph. C: control, DIE: deep infiltrating endometriosis.

Article Snippet: The CXCL1 serum levels were measured using the commercially available human CXCL1/GRO alpha Quantikine ELISA Kit in concentrations as pg/mL (Cat. number DGR00 Quantikine ELISA Human CXCL1/GRO α Immunoassay, R&D Systems).

Techniques: Comparison

Journal: International Journal of Biological Sciences

Article Title: Cancer-associated adipocytes promote the invasion and metastasis in breast cancer through LIF/CXCLs positive feedback loop

doi: 10.7150/ijbs.65227

Figure Lengend Snippet: CXCLs activate the ERK1/2 signaling pathway to up-regulate LIF expression in CAAs. ( A and B ) SB225002 or Reparixin was added to the co-culture system. ( A ) The total RNA of adipocytes was extracted for q-PCR analysis, or ( B ) the CAA culture medium was collected for ELISA analysis. ( C ) The protein expression level of adipocytes was detected by western blot. ( D ) Adipocytes were treated with rhCXCL3 or rhCXCL8 in combination with SB225002 and Reparixin. The LIF mRNA levels in adipocytes of each group were detected by q-PCR. ( E ) Adipocytes were treated with rhCXCL3 and rhCXCL8. The expression and localization of p-ERK1/2, NF-κB p65 and Stat3 in CAAs were observed using a fluorescent confocal microscope, scale bar: 20 μm. ( F and G ) α-CXCL3 (5 μg/mL) was added in the co-cultured system. The adipocyte RNA was extracted for q-PCR analysis ( F ), or the CAA culture medium was collected for ELISA analysis ( G ). ( H ) The protein expression level of adipocytes was analyzed by western blot. ( I ) The mRNA level of CXCR2 and LIF in adipocytes next to the normal breast tissue and breast cancer tissue were detected by q-PCR. ( J ) The mRNA level of CXCL1-3 and CXCL8 in normal breast tissue and breast cancer tissue was detected by q-PCR. Typical microscopic fields and blots are shown and quantitative data are presented as the mean ± SD from at least three independent experiments. # p < 0.05, ## p < 0.01, ### p < 0.001, Co-culture or rhCXCL3/8 VS. Control; * p < 0.05, ** p < 0.01, *** p < 0.001, Co-culture + inhibitors/antibody or rhCXCL3/8 + inhibitors VS. Co-culture or rhCXCL3/8.

Article Snippet: According to the manufacturer's protocol, LIF, CXCL1, CXCL2, CXCL3, and CXCL8 were measured with ELISA kits (#EK1107, #EK196, #EK1264, #EK1265, and #EK108, MultiSciences).

Techniques: Expressing, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Microscopy, Cell Culture, Control

Journal: Frontiers in Pharmacology

Article Title: Cisplatin Promotes the Efficacy of Immune Checkpoint Inhibitor Therapy by Inducing Ferroptosis and Activating Neutrophils

doi: 10.3389/fphar.2022.870178

Figure Lengend Snippet: N1-polarized neutrophils induced by cisplatin-mediated tumor ferroptosis exerted an anti-tumor effect. (A) Relative mRNA expression of CXCL1 and CXCL2 was tested by RT-qPCR assay in A549 cells treated with 5 μM DDP or rescued by 10 μM ferrostatin-1. (B) and (C) Concentration of CXCL1 (B) and CXCL2 (C) released by A549 cells in culture medium, which was tested by ELISA assay. A549 cell were treated with 5 μM DDP or rescued by 10 μM ferrostatin-1. (D) and (E) Relative mRNA expression of neutrophil cytotoxic markers (D) and pro-inflammatory markers (E) were tested by RT-qPCR assays in neutrophils co-cultured with A549 cells, which were pretreated with 5 μM DDP or rescued by ferrostatin-1. (F) Cell death ratio of A549 cells treated with DDP or rescued by ferrostatin-1 before and after being co-cultured with neutrophils. (G) Quantification of data in (F) . DDP pretreatment remarkably increases the cytotoxic effect of neutrophils which would be rescued by ferrostatin-1. Bar graphs represent the mean ± SD of indicated samples. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: ELISA was performed using human interferon gamma (IFN-γ) (430107; BioLegend, San Diego, CA, United States), human C-X-C Motif Chemokine Ligand 1 (CXCL1) (RAB0116; Sigma-Aldrich, St. Louis, MO, United States), and human C-X-C motif chemokine ligand 2 (CXCL2) (DY276-05; R&D Systems, Minneapolis, MN, United States), according to the manufacturers’ protocols.

Techniques: Expressing, Quantitative RT-PCR, Concentration Assay, Enzyme-linked Immunosorbent Assay, Cell Culture