Journal: Cancer Research

Article Title: The BH3 Mimetic ABT-737 Induces Cancer Cell Senescence

doi: 10.1158/0008-5472.can-10-1977

Figure Lengend Snippet: Figure 1. Gene transcription changes after ABT-737 treatment. A, microarray analysis of gene expression induced by ABT-737. PV-10 cells were treated in triplicate with DMSO or 10 mmol/L ABT-737 for 24 hours and gene microarray changes documented. B, gene changes associated with senescence. C, qT-PCR analysis of IL-6 and IL-8 transcripts. PV-10 and 22Rv1 cells were treated in triplicate with DMSO, ABT-737 (10 mmol/L) or enantiomer (En., 10 mmol/ L) for 24 hours (mean SD, n ¼ 3). D, the secretion of IL-6 and IL-8 in PV-10 cells treated with DMSO, ABT-737 or enantiomer for 24 hours was determined by ELISA (n ¼ 6, mean SD).

Article Snippet: Conditioned medium was collected following treatment and used at a 2:1 dilution in the human interleukin-6 (IL-6) or IL-8 ELISA Kit (R&D Systems) according to the manufacturer's instructions.

Techniques: Microarray, Gene Expression, Enzyme-linked Immunosorbent Assay

Journal: Journal of Inflammation Research

Article Title: Protective Effects of Qingre Sanjie Jiaonang on Pulmonary Fibrosis: A Pilot Study

doi: 10.2147/JIR.S479432

Figure Lengend Snippet: QRSJ alleviated the release of inflammatory factors during the progression of PF. ( A ) Immunofluorescence staining was performed, with imaging for NE (red), and staining with DAPI for nucleus (blue). Scale bar, 50 μm or 20 μm (Zoom). ( B ) Quantitative analysis of the mean fluorescence intensity of NE (n = 3). ( C ) The effect of QRSJ on the levels of NE in serum in different groups using ELISA (n = 5). ( D-F ) The effect of QRSJ on the levels of CXCL15, IL-1β and TNF-α in BALF in different groups using ELISA (n = 3). ( G ) The effect of QRSJ on the levels of IL-6 in serum in different groups using ELISA (n = 5). ( H ) The effect of QRSJ on the levels of CXCL15 in serum in different groups using ELISA (n = 4). ( I ) The effect of QRSJ on the levels of TNF-α in serum in different groups using ELISA (n = 5). The data is presented as mean ± SD, * p < 0.05, ** p < 0.01 and *** p < 0.001 by one-way ANOVA. ns represents no significant difference.

Article Snippet: Enzyme-linked immunosorbent assay (ELISA) Kits for tumor necrosis factor (TNF)-α (Cat # E-EL-M3063), IL-6 (Cat # E-EL-M0044), CXCL15 (mouse homologue of human IL-8, Cat # E-EL-M0269), HMGB1 (Cat # E-EL-M0676), IL-1β (Cat # E-EL-M0037) and NE (Cat # E-EL-M3025) were purchased from Elabscience (Wuhan, China).

Techniques: Immunofluorescence, Staining, Imaging, Fluorescence, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Veterinary Medical Science

Article Title: Effect of fuzapladib sodium hydrate on adhesion molecule expression in canine endothelial cells and neutrophils

doi: 10.1292/jvms.25-0507

Figure Lengend Snippet: Effects of fuzapladib sodium hydrate (FZP) on C-X-C motif chemokine ligand 1 (CXCL1) protein expression in cytokine-induced canine aortic endothelial cells (CnAOEC). CnAOEC were stimulated with (A) lipopolysaccharide (LPS; 100 ng/mL) or (B) recombinant canine tumor necrosis factor alpha (rh-cTNFα; 30 ng/mL), either in isolation or combined with FZP (1, 10, or 100 μM) for 24 hr. CXCL1 levels in culture supernatants were measured using a Canine CXCL1 ELISA kit. Data represent mean ± SD from three independent experiments. Statistical significance was determined by RM one-way ANOVA with Tukey’s multiple comparison test ( P >0.05).

Article Snippet: After 24 hr of stimulation, the concentration of secreted C-X-C motif chemokine ligand 1 (CXCL1) protein in the culture medium was quantified using the Canine CXCL1 ELISA kit (RayBiotech, Inc., Norcross, GA, USA) according to the manufacturer’s instructions.

Techniques: Expressing, Recombinant, Isolation, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Journal for Immunotherapy of Cancer

Article Title: Natural lung-tropic TH9 cells: a sharp weapon for established lung metastases

doi: 10.1136/jitc-2024-009629

Figure Lengend Snippet: The CXCR4-CXCL12 axis mediates the lung tropism of T H 9 cells. ( A ) The expression of chemokines in normal lung tissue was analyzed from the GSE179554 data set. ( B ) Heat map of chemokine receptor genes in T H 0 and T H 9 cells. ( C ) Real-time PCR analysis of the expression of the chemokine receptors in T H 0 and T H 9 cells. ( D ) Bubble plot depicting ranked chemokine–chemokine receptors by the production of chemokines in lungs and chemokine receptors in T H 9 cells. ( E and F ) The representative density plot ( E ) and quantification ( F ) of CD45.1-T H 9 cells in the lungs 48 hours after intravenous injection of CD45.1-T H 9 cells into healthy CD45.2 mice. Healthy CD45.2 mice were received intraperitoneal (i.p.) injection of 1 mg kg −1 ML339, C-021, R243 and AMD3100, 24 hours before T cell transfer. ( G ) Migration index of T H 1, T H 9 and T H 17 cells with the presence or absence of lung lysate (100 µg/mL). The T H cell subsets were pretreated with AMD3100 (5 µg/mL) or not. ( H ) Migration index of T H 1, T H 9 and T H 17 cells toward recombinant CXCL12 (10 ng/mL). ( I ) Immunofluorescence staining of CXCR4 in T H 1, T H 9 and T H 17 cells. Scale bar, 15 µm. ( J ) Statistical analysis of mean fluorescence intensity of CXCR4 in T H 1, T H 9 and T H 17 cells. ( K ) The intensity of CXCR4 expression detected in T H 0, T H 1, T H 9 and T H 17 cells by flow cytometry. ( L ) Western blotting analysis of CXCR4 protein levels in T H 0, T H 1, T H 9 and T H 17 cells. ( M and N ) The representative density plot ( M ) and statistical analysis ( N ) of CD45.2-T H 9 cells in the lung 48 hours after intravenous injection of CD45.2 WT derived T H 9 cells and CD45.2- Cd4 cre Cxcr4 flox/flox derived T H 9 cells into healthy CD45.1 mice. ( O ) H&E staining of lungs bearing with K7M2-OVA tumor on day 14. The mice were intravenously injected with T H 9 cells or AMD3100 (5 µg/mL) pretreated T H 9 cells on day 5 and day 12. Scale bar, 2.5 mm. ( P ) Statistical analysis of tumor weight and tumor area corresponds to ( O ). Data were analyzed by one-way ANOVA test or unpaired t-test. Representative results from three independent experiments are shown (mean±SEM); n=3 in ( C, E–H and M–P ), n=4 or 6 in ( I and J ). ns denote no significant difference,*indicates p<0.05, **indicates p<0.01, ***indicates p<0.001, ****indicates p<0.0001. ANOVA, analysis of variance; DAPI, 4'6-diamidino-2-phenylindole; OVA, ovalbumin; PBS, phosphate-buffered saline; T H , T helper; WT, wild-type.

Article Snippet: ELISA kits for murine CXCL1 (RK0038), CXCL12 (RK00168), CXCL14 (RK02643), CXCL15 (RK07476) and CXCL16 (RK00417) were all purchased from ABclonal (Wuhan, Hubei, China).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Injection, Migration, Recombinant, Immunofluorescence, Staining, Fluorescence, Flow Cytometry, Western Blot, Derivative Assay, Saline

Journal: Journal for Immunotherapy of Cancer

Article Title: Natural lung-tropic TH9 cells: a sharp weapon for established lung metastases

doi: 10.1136/jitc-2024-009629

Figure Lengend Snippet: Working model of the mechanism of T H 9 cells’ lung tropism and their antitumor effects. T H 9 cells upregulate TRAF6 expression to activate the NF-κB signaling pathway, which requires phosphorylation of IKKs and resulting in ITCH phosphorylation. Phosphorylation of ITCH by IKKs inhibits its E3 ligase activity, thereby attenuating CXCR4 degradation and ultimately resulting in its accumulation in T H 9 cells. Therefore, T H 9 cells preferentially accumulate in the lungs due to CXCL12 chemoattraction and exhibit lung tropism. In the setting of lung metastasis, transfusion of T H 9 cells inhibits tumor progression by enhancing the number of CD8 + T cells and ameliorating the immunosuppressive microenvironment with decreased TAM and MDSC. IKK, inhibitor of nuclear factor kappa B kinase; MDSC, myeloid-derived suppressor cell; TAM, tumor-associated macrophage; T H , T helper; TRAF6, tumor necrosis factor receptor associated factor 6.

Article Snippet: ELISA kits for murine CXCL1 (RK0038), CXCL12 (RK00168), CXCL14 (RK02643), CXCL15 (RK07476) and CXCL16 (RK00417) were all purchased from ABclonal (Wuhan, Hubei, China).

Techniques: Expressing, Phospho-proteomics, Activity Assay, Derivative Assay