cxcl11  (R&D Systems)

Journal: Cell Death & Disease

Article Title: Human BCL-G regulates secretion of inflammatory chemokines but is dispensable for induction of apoptosis by IFN-γ and TNF-α in intestinal epithelial cells

doi: 10.1038/s41419-020-2263-0

Figure Lengend Snippet: a – c HT-29 cells were stimulated with IFN-γ+TNF-α (1 or 10 ng/ml each) for 8 and 24 h. a Concentration of CXCL9, CXCL10, and CXCL11 measured in cell culture supernatants. b Concentration of CCL5 and CCL20 measured in cell culture supernatants. c Western blot showing levels of BCL-G L in HT-29 cells treated as indicated. d , e HT-29 cells transfected with a non-targeting siRNA (siCtrl) or siRNA targeting BCL-G (siBCL-G) were stimulated 48 h later with IFN-γ+TNF-α (1 or 10 ng/ml each) for 24 h. d Concentration of CXCL9, CXCL10 and CXCL11 measured in cell culture supernatants following BCL-G knockdown. e Concentration of CCL5 and CCL20 measured in cell culture supernatants following BCL-G knockdown. f Wild-type or BCL-G knockout (BCL-G KO ) HT-29 cells were treated with IFN-γ+TNF-α (10 ng/ml each) for 24 h. Concentration of CCL5 and CCL20 measured in cell culture supernatants (left). Luminescent Cell-Titer Glo signal of wild-type and BCL-G KO HT-29 cells after 24 h culture (right). Data shown are the mean ± S.E.M. of at least n = 3 independent experiments. * p < 0.05, ** p < 0.01 and *** p < 0.001 (two-way ANOVA followed by Tukey’s multiple comparisons test as indicated). NT — non-treated, wt — wild-type.

Article Snippet: Secreted chemokines were measured according to manufacturer’s instructions using the following DuoSet ELISA assays (Bio-Techne): CCL5 (cat# DY278), CCL20 (cat# DY360), CXCL9 (cat# DY392), CXCL10 (cat# DY266) and CXCL11 (cat# DY672).

Techniques: Concentration Assay, Cell Culture, Western Blot, Transfection, Knock-Out

Journal: Frontiers in Pharmacology

Article Title: Patient Derived Colonoids as Drug Testing Platforms–Critical Importance of Oxygen Concentration

doi: 10.3389/fphar.2021.679741

Figure Lengend Snippet: Chemokines released by colonoids at 20 or 2% O 2 . Conditioned medium from colonoids at 20 or 2% O 2 , followed by 24h treatment with TNF, IL17 or TNF/IL17, and untreated colonoids were analyzed for chemokine concentration (A) Mean protein concentration ( p g . ml −1 ) of chemokines in conditioned medium from colonoids ( n = 3 independent assays) analyzed by Human Chemokine Panel featuring 40 magnetic bead-based immunoassays. The 18 heatmaps show expression of significantly regulated proteins and are grouped according to 1) Highest protein expression by TNF/IL17 treatment at 20% O 2 . 2) Highest protein expression by TNF/IL17 treatment at 2% O 2 . 3) Highest protein expression by TNF treatment at 20% O 2 , and 4) Highest protein expression by TNF or TNF/IL17 treatment, similar at 20 and 2% O 2 (B) CCL20, CXCL1, CXCL2, CXCL5, CXCL8, CXCL10 and CXCL11 protein concentrations ( p g.ml −1 ) in conditioned medium from minimum six assays measured by ELISA. Left panels: paired data at 2 or 20% O 2 across all treatments for the selected chemokines, analyzed with Wilcoxon matched-pairs signed rank test. Significance level is indicated as p values or not significant (ns). Right panels: concentrations for each chemokine in 2% (blue) or 20% (red) O 2 plotted as individual values with mean and SD. Statistical analysis were performed on log2 transformed data . Alterations across different treatments within each oxygen concentrations were analyzed with one-way ANOVA followed by Tukey’s multiple comparisons or Kruskal-Wallis test followed by Dunn’s multiple comparisons test. Alterations across different oxygen concentrations within same treatment were analyzed by two-way ANOVA followed by Holm-Šídák multiple comparisons test. * p < 0.05; red and blue asterisks show comparisons across different treatments within 20 and 2% O 2 conditions, respectively. Black asterisks indicate significant comparisons between 2 and 20% oxygen concentrations within a specific treatment.

Article Snippet: ELISA kits for human CXCL1 (DY275-05), CXCL2 (DY276-05), CXCL5 (DY254-05), CXCL8 (DY208), CXCL10 (DY266-05), CXCL11 (DY672) and CCL20 (DY360), all from R&D Systems (Abington, United Kingdom), were used according to manufacturer’s protocols.

Techniques: Concentration Assay, Protein Concentration, Expressing, Enzyme-linked Immunosorbent Assay, Transformation Assay