Journal: Brazilian Journal of Cardiovascular Surgery

Article Title: Expression Level and Significance of Tim-3 in CD4 + T Lymphocytes in Peripheral Blood of Patients with Coronary Heart Disease

doi: 10.21470/1678-9741-2020-0509

Figure Lengend Snippet: Upregulation of the expression of IL-7 in peripheral blood serum of patients with CHD. The expression level of IL-7 in peripheral blood serum of the study subjects in each group was detected by ELISA. *Compared with the control group, *P<0.05, **P<0.01; # P<0.05.

Article Snippet: The level of IL-7 in peripheral blood serum was detected using an IL-7 ELISA kit (SEKH-0015, Solarbio, Beijing, China) in strict accordance with the instructions of the kit.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Brazilian Journal of Cardiovascular Surgery

Article Title: Expression Level and Significance of Tim-3 in CD4 + T Lymphocytes in Peripheral Blood of Patients with Coronary Heart Disease

doi: 10.21470/1678-9741-2020-0509

Figure Lengend Snippet:

Article Snippet: The level of IL-7 in peripheral blood serum was detected using an IL-7 ELISA kit (SEKH-0015, Solarbio, Beijing, China) in strict accordance with the instructions of the kit.

Techniques: Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction, Reverse Transcription, Standard Deviation

Journal: The Journal of Experimental Medicine

Article Title: Dietary restriction improves repopulation but impairs lymphoid differentiation capacity of hematopoietic stem cells in early aging

doi: 10.1084/jem.20151100

Figure Lengend Snippet: IGF1 injection reverts the increases in HSC quiescence but not the suppression of lymphopoiesis in mice exposed to DR. (A) ELISA-determined serum level of IGF1 in mice treated with DR or AL for 4 d or refed with an AL diet for 3 d after 4 d of DR ( n = 3–4 mice per group; n = 2 independent experiments). (B) ELISA-determined serum level of IGF1 in 2-mo-old p53 −/− mice and p53 +/+ mice (littermates) that were treated with DR or AL for 2 wk ( n = 3–4 mice per group; n = 2 independent experiments). (C–G) Mice were treated with a DR or AL diet for 8 d. In parallel, recombinant human IGF1 protein or vehicle control was subcutaneously injected twice per day at a dose of 500 µg/kg ( n = 4 mice per group; n = 2 independent experiments). FACS analysis was performed 24 h after the last injection on freshly isolated BM cells: percentage of HSCs in the indicated cell cycle stages (C), total numbers of CLPs (D), pro–B cells (E), MEPs (F), and proEry’s (G) per million total BM cells are show. AL con, control-injected AL mice; AL IGF1, IGF1-injected AL mice; DR con, control-injected DR mice; DR IGF1, IGF1-injected DR mice. Data are displayed as mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 by one-way ANOVA. ns, not significant.

Article Snippet: ELISA for detecting IGF1, IL-6, IL-7, and total corticosterone was performed using the IGF-I mouse/rat ELISA kit (Mediagnost), the Mouse IL-6 Quantikine ELISA kit (R&D Systems), the Mouse IL-7 Quantikine ELISA kit (R&D Systems), and Corticosterone EIA kit (ARBOR ASSAYS), respectively, according to the manufacturer’s instructions.

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Recombinant, Control, Isolation

Journal: The Journal of Experimental Medicine

Article Title: Dietary restriction improves repopulation but impairs lymphoid differentiation capacity of hematopoietic stem cells in early aging

doi: 10.1084/jem.20151100

Figure Lengend Snippet: IL-7 or IL-6 injection reverts suppression of lymphopoiesis in mice exposed to DR. (A) ELISA-determined serum level of IL-7 in mice treated with DR or AL for 7 d or refed with AL diet for 3 d after 7 d of DR ( n = 3–4 mice per group; n = 2 independent experiments). One-way ANOVA. (B–G) 2-mo-old mice were treated with DR or AL diet for 9 d. In parallel, mouse IL-7 protein or vehicle control was subcutaneously injected for 9 d at a dose of 50 µg/kg once per day ( n = 5 mice per group; n = 2 independent experiments). Mice were sacrificed for analysis 24 h after the last injection. (B–D) FACS analysis was performed on freshly isolated BM cells to determine the total number of CLPs (B) and MEPs (C), and the percentage of CLPs (D) of DR mice in the indicated phases of the cell cycle (FACS analysis by Ki67 and DAPI staining). (E and F) the ratio of CLPs versus lymphoid-biased HSCs (E) and of pro–B cells versus CLPs (F). (G) The mRNA expression levels of genes that are associated with lymphoid cell differentiation and erythroid cell differentiation were determined by qRT-PCR in freshly isolated lymphoid-biased HSCs under the indicated treatment conditions. One-way ANOVA (B, C, and E–G) or unpaired two-tailed Student’s t test (D) was used. (H) ELISA-determined serum level of IL-6 in mice treated with DR or AL for 7 d or refed with AL diet for 3 d after 7 d of DR ( n = 3–4 mice per group; n = 2 independent experiments). One-way ANOVA was used. (I) 2-mo-old mice were treated with a DR or AL diet for 5 d. In parallel, mouse IL-6 protein or vehicle control was subcutaneously injected into mice daily at a dose of 50 µg/kg ( n = 4 mice per group; n = 2 independent experiments). The total number of CLPs was determined by FACS analysis, 24 h after the last injection. One-way ANOVA analysis. (J–P) ADX or sham surgery was performed on 2-mo-old mice. 10 d after operation, mice were treated with DR or AL for 9 d ( n = 4–5 mice per group; n = 2 independent experiments). Mice of the indicated cohorts were analyzed as follows: (J, O, and P) ELISA-determined serum levels of total corticosterone (J), IL-7 (O), and IL-6 (P); (K–M) FACS analysis of the total number of CLPs (K) and MEPs (M) per million BM cells and of the percentage of CLPs (L) of DR mice in different stages of the cell cycle (by Ki67 and DAPI staining); (N) expression of lymphoid- and erythroid-specific genes in lymphoid-biased HSCs of mice of the indicated cohorts. One-way ANOVA (J, K, and M–P) or unpaired two-tailed Student’s t test (L) was used. AL con, control-injected AL mice or sham-operated AL mice; AL IL-7, IL-7–injected AL mice; DR con, control-injected DR mice or sham-operated DR mice; DR IL-7, IL-7–injected DR mice; AL IL-6, IL-6–injected AL mice; DR IL-6, IL-6–injected DR mice; AL ADX, AL mice with ADX; DR ADX, DR mice with ADX. Data are displayed as mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. ns, not significant.

Article Snippet: ELISA for detecting IGF1, IL-6, IL-7, and total corticosterone was performed using the IGF-I mouse/rat ELISA kit (Mediagnost), the Mouse IL-6 Quantikine ELISA kit (R&D Systems), the Mouse IL-7 Quantikine ELISA kit (R&D Systems), and Corticosterone EIA kit (ARBOR ASSAYS), respectively, according to the manufacturer’s instructions.

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Control, Isolation, Staining, Expressing, Cell Differentiation, Quantitative RT-PCR, Two Tailed Test

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: BAG2 Inhibits Cervical Cancer Progression by Modulating Type I Interferon Signaling through Stabilizing STING.

doi: 10.1002/advs.202414637

Figure Lengend Snippet: Figure 6. BAG2 positively regulates STING-mediated type I interferon responses. A) RNA seq data of SiHa cells after BAG2 overexpression were analyzed by gene set enrichment analysis (GSEA), and Hallmark gene sets were used as annotated gene sets. B) Gene heat map of Hallmark Interferon 𝛼Response gene set after BAG2 overexpression in SiHa cells. C) SiHa cells were transfected with vector or GFP-BAG2 for 48 h, and then they were treated with HT- DNA for 0, 3, 6 h or cGAMP for 0, 2, 4 h. Cells were collected for detection of STING, p-TBK1, TBK1, p-IRF3, and IRF3 protein levels by Western blot. D) Vector or BAG2 plasmid was transfected in SiHa cells for 48 h, cell supernatants were collected by HT-DNA treatment for 6 hrs, and IFN-𝛽and IL7 production was measured by ELISA. E) Vector or BAG2 plasmid was transfected in U14 cells for 48 h, cell supernatants were collected by HT-DNA treatment for 6 h, and IFN-𝛽and IL7 production was measured by ELISA. F) SiHa cells and G) U14 cells were transfected with Vector or BAG2 for 48 h, then cGAMP was added for 4 h. qRT-PCR was used to measure the mRNA expression of ISG15, IFN-𝛽, MX1, and CXCL10. Data are presented as mean values ± SD, with n = 3 (B, D-G) biological independent experiments. Statistical significance was determined by two-tailed unpaired Student’s t-test (D-G).

Article Snippet: ELISA Assay: After transfecting the cells, the supernatant was collected, centrifuged to remove the precipitate, and the samples were processed according to the instructions of IL7 kit (EK207, EK107, MultiSciences Biotech) and IFN-β kit (EK2236, EK1236,MultiSciences Biotech), and then the absorbance value was detected by a microplate reader.

Techniques: RNA Sequencing, Over Expression, Transfection, Plasmid Preparation, Western Blot, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing, Two Tailed Test

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Soluble IL7R? potentiates IL-7 bioactivity and promotes autoimmunity

doi: 10.1073/pnas.1222303110

Figure Lengend Snippet: sIL7Rα binds IL-7 but not TSLP. The binding kinetics for all of the cytokine/receptor interactions fit best to a two-step binding reaction model using two on-rate (k1 and k2) and off-rate (k-1 and k-2) constants. (A–D) Surface plasmon resonance sensorgrams are shown for each designated protein pair. Black lines represent raw data and red lines represent the global fitting analysis of the sensorgrams to a two-step binding reaction model using ClampXP. (E) Summary of binding affinities measured in A–D. Experiments were performed in triplicate with errors on the order of 5–10% for the rate constants.

Article Snippet: Human IL-7 levels in human plasma, mouse plasma, and culture supernatants were measured by high-sensitivity IL-7 ELISA kit according to the manufacturer’s instructions (Quantikine; R&D Systems).

Techniques: Binding Assay, SPR Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Soluble IL7R? potentiates IL-7 bioactivity and promotes autoimmunity

doi: 10.1073/pnas.1222303110

Figure Lengend Snippet: sIL7RΑ potentiates IL-7–mediated survival and diminishes consumption of IL-7. (A) Survival of the IL-7–dependent cell line 2E8 was measured in the presence of rhIL-7 (2,000 pg/mL) plus varying concentrations of sIL7Rα. No effects were seen at any time point using molar ratios of 128:1 and below. However, on day 12, increased survival was observed at sIL7Rα:IL-7 molar ratios of 256–512:1, which was diminished at higher molar ratios. (B) Using lower concentrations of rhIL-7 [250 pg/mL (Left) and 500 pg/mL (Right)], survival of 2E8 from days 0–7 is shown with no cytokine, rhIL-7 alone, or sIL7Rα:rhIL7 molar ratio of 500:1. sIL7RΑ significantly increased survival on days 5 and 7 (*P < 0.05; **P < 0.01; ***P < 0.001). Error bars represent SEM of triplicate experiments. (C) IL-7 levels measured in the cultures described in B. Significantly increased IL-7 levels were measured in the presence of sIL7Rα on days 1, 3, and 5 of culture (*P < 0.05; **P < 0.01; ***P < 0.001). Error bars represent SEM of triplicate experiments. This experiment was performed three times with similar results.

Article Snippet: Human IL-7 levels in human plasma, mouse plasma, and culture supernatants were measured by high-sensitivity IL-7 ELISA kit according to the manufacturer’s instructions (Quantikine; R&D Systems).

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Soluble IL7R? potentiates IL-7 bioactivity and promotes autoimmunity

doi: 10.1073/pnas.1222303110

Figure Lengend Snippet: sIL7Rα-mediated modulation of IL-7 signaling on human T cells leads to diminished IL-7 consumption and diminished SOCS1 and CD95 Induction. Human PBMCs were coincubated with rhIL-7 with or without sIL7Rα and then analyzed at the times shown. Molar ratios were as noted in A, whereas 500:1 sIL7Rα:rhIL7 molar ratio was used in B–D. (A) IL-7–induced STAT-5 phosphorylation was measured in gated CD4+ and CD8+ T cells after 15 min. Cells were serum-starved before IL-7 was added. No effect was seen at 50:1 molar ratio, incomplete inhibition was seen at 500:1 ratio (similar effects at 1 and 10 ng/mL rhIL-7), and complete inhibition was seen at 5,000:1 ratio. Controls using human serum albumin and rat IgG2a at similar concentrations showed no significant impact on STAT-5 signaling. (B) Modulation of IL-7–induced changes in CD127, CXCR4, and CD95 expression by sIL7Rα. Representative flow-cytometric histograms of CD127 expression at day 5 on CD4+ and CD8+ T cells are shown on the right. (C) IL-7 consumption by human PBMCs is diminished in the presence of sIL7Rα, as measured by ELISA. (D) SOCS1 levels are reduced in the presence of sIL7Rα after 24 h of incubation with IL-7. Relative quantity (RQ) of SOCS1/GAPDH mRNA expression compared with untreated cells was determined. Error bars represent 95% CI of triplicate experiments. A total of three independent experiments were performed using PBMCs from different donors with comparable results. Statistical significance shown (*P < 0.05) reflects comparisons between rhIL-7 alone and rhIL-7 plus sIL7Rα using a two-tailed t test.

Article Snippet: Human IL-7 levels in human plasma, mouse plasma, and culture supernatants were measured by high-sensitivity IL-7 ELISA kit according to the manufacturer’s instructions (Quantikine; R&D Systems).

Techniques: Inhibition, Expressing, Enzyme-linked Immunosorbent Assay, Incubation, Two Tailed Test

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Soluble IL7R? potentiates IL-7 bioactivity and promotes autoimmunity

doi: 10.1073/pnas.1222303110

Figure Lengend Snippet: sIL7Rα diminishes IL-7 clearance in vivo and increases IL-7–mediated homeostatic peripheral expansion. (A) On day 0, IL-7−/− mice received one dose of rhIL-7 (5 μg) with or without sIL7Rα (100 μg) coinjected in the same syringe. A 10:1 molar ratio was chosen because of limited availability of recombinant sIL7Rα. Plasma levels were measured at 24 and 96 h. Mice injected with rhIL-7 plus sIL7Rα have higher plasma IL-7 levels after 24 h than those receiving rhIL-7 alone. (B) On day 0, mice received IL-7 with or without sIL7Rα, as in A above, plus 2 × 106 congenic lymph node cells. On day 8, mice injected with rhIL-7 plus sIL7Rα showed higher numbers of adoptively transferred CD4+ and CD8+ splenocytes compared with mice receiving rhIL-7 alone. Asterisks denote significant differences. These experiments were repeated once for two independent experiments with similar results (n = 5 per group). (C) C57BL/6 mice (n = 10/group) were immunized with MOG and then scored for EAE symptoms. PBS, rhIL-7, sIL7Rα, or rhIL-7 plus sIL7Rα was administered i.p. on days 9 and 15 postimmunization (arrows). Mice injected with rhIL-7 plus sIL7Rα showed increased EAE scores (Left), more rapid progression to EAE score 3 (Center), and higher overall disability as measured by area under the curve of EAE score over time per individual mouse (Right). *P < 0.05 as determined by Mann–Whitney test. No significant differences were seen when injecting IL-7 with a control protein (αhIL6Rα) compared with IL-7 alone, using the models shown in A, B, or C. This experiment was carried out three times with similar results.

Article Snippet: Human IL-7 levels in human plasma, mouse plasma, and culture supernatants were measured by high-sensitivity IL-7 ELISA kit according to the manufacturer’s instructions (Quantikine; R&D Systems).

Techniques: In Vivo, Recombinant, Injection, MANN-WHITNEY

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Soluble IL7R? potentiates IL-7 bioactivity and promotes autoimmunity

doi: 10.1073/pnas.1222303110

Figure Lengend Snippet: IL7R*CC MS patients have increased plasma IL-7 levels. (A) IL-7 levels were measured in plasma and CSF from patients with MS according to genotype. IL7R*CC patients had significantly higher mean plasma IL-7 levels than IL7R*TT patients, whereas no difference was observed between IL7R*CC and IL7R*CT heterozygotes. No correlation between CSF IL-7 levels and IL7R genotype was observed. The dotted line denotes limit of detection. (B) Plasma IL-7 levels were measured in plasma from a second independent cohort of patients with MS, as well as a separate cohort of healthy controls. IL7R*CC MS patients have significantly higher mean IL-7 levels than IL7R*TT MS patients, whereas no difference was observed according to genotype in healthy controls. Each shape represents a plasma or CSF sample analyzed from one patient or healthy control.

Article Snippet: Human IL-7 levels in human plasma, mouse plasma, and culture supernatants were measured by high-sensitivity IL-7 ELISA kit according to the manufacturer’s instructions (Quantikine; R&D Systems).

Techniques:

Journal: Journal of autoimmunity

Article Title: Interleukin-7 and -15 Maintain Pathogenic Memory Th17 Cells in Autoimmunity

doi: 10.1016/j.jaut.2016.11.003

Figure Lengend Snippet: (A) Compared to the normal control (NL), there was a significant memory Th17 population (CD44hiIL-17+CD4+) at both the inflamed eye (conjunctivae, CONJ) and the draining lymph nodes (DLNs) in autoimmune dry eye disease (DED). Representative flow cytometry dot plots are shown on the left. Data presented on bar graphs are combined results from three independent experiments (n = 5 mice / experiment) and represent mean±SEM. (B) Representative histographs showing IL-7Rα and IL-15Rα expression (blue lines) on memory Th17 cells from DED mice. Gray areas represent isotype controls. IL-15Rα expression on memory Th17 is highly variable. (C) mRNA and protein expression of IL-7 in CONJ were quantified by real-time RT-PCR and ELISA, respectively, and mRNA data were normalized to normal mice (NL). (D) mRNA and protein expression of IL-15 in DLN were quantified by real-time RT-PCR and ELISA, respectively, and mRNA data were normalized to normal mice (NL). Data shown in (C) and (D) are combined from two independent experiments (n = 5 mice / experiment) and represent mean±SEM. *, p < 0.05.

Article Snippet: The supernatant was assayed using commercial ELISA kits for levels of the total protein (Thermo Scientific), IL-7, and IL-15 (eBioscience).

Techniques: Flow Cytometry, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Journal of autoimmunity

Article Title: Interleukin-7 and -15 Maintain Pathogenic Memory Th17 Cells in Autoimmunity

doi: 10.1016/j.jaut.2016.11.003

Figure Lengend Snippet: (A) Whole and intact DLNs were collected from autoimmune DED and cultured in complete RPMI-1640 with anti-IL-7, anti-IL-7 and exogenous IL-15, anti-IL-15, anti-IL-15 and exogenous IL-7, anti-IL-7 and anti-IL-15 Abs, exogenous IL-7, IL-15, or IL-7 and IL-15 for 72 hours. Thereafter, the DLNs were analyzed for memory Th17 cell (mTh17) frequencies by flow cytometry. Representative flow cytometry graphs are shown on the bottom. Data presented on the top bar graphs are normalized to medium only group and combined from two independent experiments (n = 4–5 wells in each group in each experiment) and represent mean±SEM. *, decrease with p < 0.05 as compared to medium control (with isotype IgG); †, increase with p < 0.05 as compared to medium control. No significant differences were observed between anti-IL-7 and anti-IL-7 + IL-15, between anti-IL-15 and anti-IL-15 + IL-7, or between IL-7, IL-15, and IL-7 + IL-15 groups. (B) CONJ were collected from DED mice and cultured in complete RPMI-1640 with anti-IL-7, anti-IL-15, or anti-IL-7 and anti-IL-15 Abs for 72 hours. Thereafter, CONJ were analyzed for memory Th17 cell (mTh17) frequencies by flow cytometry. Data are representative of 2 experiments (n = 5 pooled eyes in each group).

Article Snippet: The supernatant was assayed using commercial ELISA kits for levels of the total protein (Thermo Scientific), IL-7, and IL-15 (eBioscience).

Techniques: Cell Culture, Flow Cytometry

Journal: Journal of autoimmunity

Article Title: Interleukin-7 and -15 Maintain Pathogenic Memory Th17 Cells in Autoimmunity

doi: 10.1016/j.jaut.2016.11.003

Figure Lengend Snippet: Memory CD4+ T cells (CD44hiCD62L−CD4+) were isolated from the draining lymph nodes of DED mice and then cultured in the presence of different cytokines, Abs, and signaling inhibitors as listed in the table. After 72 hours, the cells were collected and analyzed by flow cytometry. Representative flow graphs (top) and bar graphs (bottom) show frequencies of memory Th17 cell (A), frequencies of proliferating Ki-67+ memory Th17 cell (B), and frequencies of Annexin V+ memory Th17 cell (C). Data represent mean±SEM (n = 4 wells in each group). *, p < 0.05 as compared to IL-7+IL-15 group.

Article Snippet: The supernatant was assayed using commercial ELISA kits for levels of the total protein (Thermo Scientific), IL-7, and IL-15 (eBioscience).

Techniques: Isolation, Cell Culture, Flow Cytometry

Journal: Journal of autoimmunity

Article Title: Interleukin-7 and -15 Maintain Pathogenic Memory Th17 Cells in Autoimmunity

doi: 10.1016/j.jaut.2016.11.003

Figure Lengend Snippet: Autoimmune DED was induced on day 0 and treated with anti-IL-7, anti-IL-15, anti-IL-17 Ab, or control IgG from day 14 to 28. (A) Clinical disease severity was evaluated by corneal fluorescein staining with the representative images exhibited. The summary data shown represent mean±SEM from one representative experiment out of two performed (n = 10–14 eyes in each group). *, p < 0.05 as compared to IgG control. (B) Frequencies of memory Th17 cells (mTh17) at both CONJ and DLN were assessed by flow cytometry. *, p < 0.05 as compared to IgG control; †, p < 0.05 as compared to anti-IL-17 Ab. (C) At the end of the treatment (day 28), all groups were re-challenged with desiccating stress for 8 days. Naïve mice served as a primary challenge control. Ocular disease severity was evaluated by corneal fluorescein staining with the disease score changes calculated as Δ epithelial disease score = disease score after re-challenge − disease score before re-challenge (left) and the representative images shown (right). Summary data represent mean±SEM from 6–8 eyes. Conjunctivae tissues were analyzed for Th17 cell frequencies by flow cytometry after 8 days of re-challenge. *, p < 0.05 as compared to non-pre-treated, new onset DED group; †, p < 0.05 as compared to anti-IL-17 Ab pre-treatment.

Article Snippet: The supernatant was assayed using commercial ELISA kits for levels of the total protein (Thermo Scientific), IL-7, and IL-15 (eBioscience).

Techniques: Staining, Flow Cytometry

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: TGFß1 Stimulates Lymphatic Endothelial Cells to Produce IL7 and IL15, Which Act as Chemotactic Factors for Breast Cancer Cells with Mesenchymal Properties

doi: 10.1007/s10911-023-09552-y

Figure Lengend Snippet: Induction of IL7 and IL15 in SV-LEC cells by TGFβ1. ( A, B ) QPCR results validating the RNA seq data showing induction of Il7 and Il15 in TGFβ1 treated compared to non-treated SV-LEC cells. ( C, D ) Results from ELISA assays showing increased IL7 and IL15 levels in conditioned medium from SV-LEC cells exposed to TGFβ1 compared to control. Data represent mean results from three independent experiments with three technical replicates per condition and experiment. *** = P < 0.001; * = P < 0.05

Article Snippet: The cell culture medium of svLEC stimulated by the TGFβ1 (10 ng/ml) for 72 h was tested by enzyme-linked immunosorbent assay (ELISA) kit for detection of the concentration of mouse IL7 (Proteintech, KE10028) and IL5 (Proteintech, KE10060).

Techniques: RNA Sequencing, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: TGFß1 Stimulates Lymphatic Endothelial Cells to Produce IL7 and IL15, Which Act as Chemotactic Factors for Breast Cancer Cells with Mesenchymal Properties

doi: 10.1007/s10911-023-09552-y

Figure Lengend Snippet: IL7 and IL15 act as chemoattractants for breast cancer cells with mesenchymal properties. ( A ) Results from invasion assays showing the capacity of IL7 (10 ng/ml) and IL15 (10 ng/ml) to promote migration of NMuMG cells pre-treated or not treated with TGFβ1. ( B, C ) Results from invasion assays showing that IL7 and IL15 ( B ) as well as TNFa (10 ng/ml) ( C ) stimulate migration of MDA-MB-231 cells more than control (0.5% FBS). ( D ) Results from invasion assays showing the effect of conditioned medium (CM) from untreated (-TGFβ1) or TGFβ1-treated (10 ng/ml, 72 h) SV-LECs on the invasion of MDA-MB-231 cells in the presence of a neutralizing antibody against IL7 (IL7Ab), or an isotype control IgG antibody (IgG). Data represent mean results from three independent experiments with three technical replicates per condition and experiment. **** = P < 0.0001; *** = P < 0.001; ** = P < 0.01; * = P < 0.05

Article Snippet: The cell culture medium of svLEC stimulated by the TGFβ1 (10 ng/ml) for 72 h was tested by enzyme-linked immunosorbent assay (ELISA) kit for detection of the concentration of mouse IL7 (Proteintech, KE10028) and IL5 (Proteintech, KE10060).

Techniques: Migration, Control

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: TGFß1 Stimulates Lymphatic Endothelial Cells to Produce IL7 and IL15, Which Act as Chemotactic Factors for Breast Cancer Cells with Mesenchymal Properties

doi: 10.1007/s10911-023-09552-y

Figure Lengend Snippet: TGFβ1 promotes chemotactic migration of breast cancer cells towards lymphatics. Schematic summary of the results indicating that TGFβ1 plays dual roles in lymphatic dissemination of breast cancer cells by (i) inducing EMT and providing tumor cells with mesenchymal and invasive properties and (ii) stimulating lymphatic endothelial cells to produce chemotactic factors including IL7 and IL15. As a result, breast cancer cells with mesenchymal properties, which overexpress IL7 and IL15 receptors, are activated for chemotactic migration towards the lymphatics

Article Snippet: The cell culture medium of svLEC stimulated by the TGFβ1 (10 ng/ml) for 72 h was tested by enzyme-linked immunosorbent assay (ELISA) kit for detection of the concentration of mouse IL7 (Proteintech, KE10028) and IL5 (Proteintech, KE10060).

Techniques: Migration

Journal: Cell stem cell

Article Title: Bone marrow myeloid cells regulate myeloid-biased hematopoietic stem cells via a histamine-dependent feedback loop

doi: 10.1016/j.stem.2017.11.003

Figure Lengend Snippet: (A) TLR4 mRNA in HSCs (n = 3 - 4) and myeloid cells (n = 5). (B) TLRs expression from gene microarray analysis in myeloid cells (n = 3 per group). (C) Quantification of HSCs, HSPCs, and myeloid cells at 24 hours after LPS treatment (n = 5). (D) Myeloid cells (Gr1+) and progenitors (c-kit+) in spleen from LPS (n = 4) or PBS (n = 3)-treated Hdc-GFP mice. Four independent experiments. (E and F) Cell cycle analysis of BM HSCs at 6 hours after PBS (n = 3) or LPS (n = 3) treatment. (G) Absolute numbers of HSC in BM of Hdc-GFP mice treated with PBS, IFN-γ, or IL6 (n = 5 per group). Data were analyzed with two-tailed Student’s t-test (A, F, and G) or one-way analysis of variation (ANOVA) with Bonferroni post-hoc test (C). See also Figure S3.

Article Snippet: Proteins from serum or bone marrow supernatant were measured by using ELISA Kit (Histamine: Enzo Life Science; IL6, IL7, and IFN-γ: Thermo Fisher Scientific) according to manufacturer’s protocol.

Techniques: Expressing, Microarray, Cell Cycle Assay, Two Tailed Test

Journal: Cell stem cell

Article Title: Bone marrow myeloid cells regulate myeloid-biased hematopoietic stem cells via a histamine-dependent feedback loop

doi: 10.1016/j.stem.2017.11.003

Figure Lengend Snippet: (A) Relative mRNA expression of IL-7/IL7-R pathway genes in HSPCs (n = 4 per group). (B) Quantification of Hdc-GFPhi HSPCs in IL-7 or PBS-treated mice (n = 3 per group). (C) Percentage of G0 HSCs (n = 3 per group). (D) Frequencies of progenitors in IL-7 or PBS-treated Hdc-GFP mice (n = 5 per group). (E) Schematic depiction of the relative lineage bias of Hdc-GFPhi HSCs or Hdc-GFPlo HSCs. Data were analyzed with two-tailed Student’s t-test (A-D). See also Figure S3.

Article Snippet: Proteins from serum or bone marrow supernatant were measured by using ELISA Kit (Histamine: Enzo Life Science; IL6, IL7, and IFN-γ: Thermo Fisher Scientific) according to manufacturer’s protocol.

Techniques: Expressing, Two Tailed Test

Journal: Cell stem cell

Article Title: Bone marrow myeloid cells regulate myeloid-biased hematopoietic stem cells via a histamine-dependent feedback loop

doi: 10.1016/j.stem.2017.11.003

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Proteins from serum or bone marrow supernatant were measured by using ELISA Kit (Histamine: Enzo Life Science; IL6, IL7, and IFN-γ: Thermo Fisher Scientific) according to manufacturer’s protocol.

Techniques: Purification, Recombinant, Lysis, Staining, Enzyme-linked Immunosorbent Assay, Sequencing, Isolation, SYBR Green Assay, Expressing, Software