Journal: International immunopharmacology

Article Title: Tumor cell derived osteopontin and prostaglandin E2 synergistically promote the expansion of myeloid derived suppressor cells during the tumor immune escape phase.

doi: 10.1016/j.intimp.2024.111584

Figure Lengend Snippet: Fig. 5. Increased production of osteopontin (OPN) and prostaglandin E2 (PGE2) in EL4luc2 cells and associated tumor model. (a and b), Expression of angiogenic factors and cytokines produced by EL4luc2 cells was screened using proteome profiler kits for mouse angiogenesis factors (a) and cytokines/chemokines (b). (c-e), levels of OPN and PGE2, and IL-17 were measured in the control culture medium (white bars: c, d, e) EL4 cells-derived cell culture medium (black bars: c, d, e) and sera from tumor free (control) mice (white bars: c, d, e), and EL4luc2 tumor-bearing mice (black bars: c, d, e), respectively. Statistical analysis was performed using the Mann-Whitney U test (*P ≤0.05).

Article Snippet: The levels of PGE2, OPN and IL-17 in the cell culture supernatants and serum samples were quantified using commercially available ELISA kits (Mouse Prostaglandin E2 (PGE2) ELISA Kit, MyBioSource, Inc. San Diego, CA, USA, Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit, R&D Systems, Abingdon, UK, and ELISA Flex: Mouse IL-17A (HRP), Mabtech AB, Nacka Strand, Sweden), in accordance with the manufacturers’ instructions.

Techniques: Expressing, Produced, Control, Derivative Assay, Cell Culture, MANN-WHITNEY

Journal: International immunopharmacology

Article Title: Tumor cell derived osteopontin and prostaglandin E2 synergistically promote the expansion of myeloid derived suppressor cells during the tumor immune escape phase.

doi: 10.1016/j.intimp.2024.111584

Figure Lengend Snippet: Fig. 6. Osteopontin (OPN) and prostaglandin E2 (PGE2) synergistically induce myelopoiesis with immunosuppressive activity. (a-d) Freshly flushed bone marrow (BM) cells were cultured in a 24-well cell culture plate for 72 h in the presence of various combinations of PGE2, OPN, and IL-17. Subsequently, cell viability (a and d) and immunophenotypic analysis of myeloid cells (b and e) flow cytometry analyses were conducted on the cultured BM cells using trypan blue exclusion test and flow cytometry, respectively. Representative photographs of myeloid cell populations are displayed above respective bars (b and e). c) Conditioned medial levels of arginase were quantified using colorimetric assays after treating BM cells with PGE2 and OPN. f) Effect of pioglitazone on the inhibition of proliferation of EL4 cells. Statistical analysis was performed using ANOVA test (*P ≤0.05, **P < 0.01, ***P < 0.01).

Article Snippet: The levels of PGE2, OPN and IL-17 in the cell culture supernatants and serum samples were quantified using commercially available ELISA kits (Mouse Prostaglandin E2 (PGE2) ELISA Kit, MyBioSource, Inc. San Diego, CA, USA, Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit, R&D Systems, Abingdon, UK, and ELISA Flex: Mouse IL-17A (HRP), Mabtech AB, Nacka Strand, Sweden), in accordance with the manufacturers’ instructions.

Techniques: Activity Assay, Cell Culture, Flow Cytometry, Inhibition

Journal: Frontiers in Endocrinology

Article Title: Associations between seminal plasma osteopontin level and sperm motility in infertile men with asthenozoospermia

doi: 10.3389/fendo.2025.1487650

Figure Lengend Snippet: OPN concentration in the seminal plasma of infertile men with AZS (n=78) compared to the fertile men (n=80). (A) Scatter plots of OPN concentration in the seminal plasma of infertile men with AZS patients and fertile group using ELISA method. Data are presented as mean ± SEM; ∗∗∗∗ P < 0.0001. (B) ROC curves assessing the discriminative capability of OPN levels between infertile men with AZS and healthy fertile men, showing significant discriminative power with an AUC value of 0.793 (95% Confidence Interval, 0.724–0.863).

Article Snippet: According to the manufacturer’s instructions, seminal plasma OPN concentration was evaluated using the Human Osteopontin (OPN) Quantikine ELISA Kit (R&D systems, Minneapolis, MN, USA, catalog number DOST00).

Techniques: Concentration Assay, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Journal of Biomedical Research

Article Title: DEC1 promotes breast cancer bone metastasis through transcriptional activation of CXCR4

doi: 10.7555/JBR.39.20250031

Figure Lengend Snippet: DEC1 deficiency prevented breast bone metastasis induced by intracardiac injections of 4T1 cells via decreasing CXCR4/CXCL12 in mice. Forty 4-month-old mice, comprising twenty Dec1 +/+ and twenty Dec1 −/− , were divided into four groups: Dec1 +/+ -PBS, Dec1 +/+ -4T1, Dec1 −/− -PBS, and Dec1 −/− -4T1. Mice in the Dec1 +/+ -4T1 and Dec1 −/− -4T1 groups received an intracardiac injection of 4T1 mouse breast cancer cells to induce breast bone metastasis, while mice in the Dec1 +/+ -PBS and Dec1 −/− -PBS groups received an equal volume of PBS via the same method for two months. n = 10 for each group. A: Osteolytic bone injury in the four groups of mice by X-ray. B: The serum CXCL12 amount in the four groups of mice ( n = 3–5). C–F: The CA153, CK8, and Ki67 expression in the four groups of mice by immunohistochemical staining ( n = 3). G and H: The TRAP-positive cells in the four groups of mice ( n = 3). I–N: The protein levels of CXCR4, TRAP, N-cadherin, E-cadherin, and vimentin in the four groups of mice by Western blotting ( n = 3). O: The CXCR4 expression in the four groups of mice. Data are presented as mean ± standard deviation. * P < 0.05, ** P < 0.01, *** P < 0.001, and ns P > 0.05, comparisons are shown in the figure. Data were analyzed using two-way ANOVA, and differences between groups were analyzed using Student's t -test.

Article Snippet: CXCL12 enzyme-linked immunosorbent assay (ELISA) Kit (Cat. #KE10046) was from Proteintech (Wuhan, China).

Techniques: Injection, Expressing, Immunohistochemical staining, Staining, Western Blot, Standard Deviation

Journal: Journal of Biomedical Research

Article Title: DEC1 promotes breast cancer bone metastasis through transcriptional activation of CXCR4

doi: 10.7555/JBR.39.20250031

Figure Lengend Snippet: DEC1 promoted CXCL12 production from mesenchymal stromal cells in mice and MC3T3-E1 cells. A: The CXCL12 expression in the femur of wildtype (WT) and Dec1 -knockout (KO) mice by immunohistochemical staining ( n = 3 in each group). B: The relative Cxcl12 mRNA levels in bone tissues from the femur of WT and Dec1 -KO mice ( n = 5 in each group). C: The relative Cxcl12 mRNA levels in mesenchymal cells from the femur of WT and Dec1 -KO mice ( n = 3 in each group). D–F: MC3T3-E1 cells were seeded into 6-well plates and cultured overnight. Cells were divided into three groups: Vector (infected with LV-shCon and transfected with Flag-CMV2), DEC1 -OE (transfected with Flag- DEC1 ), and DEC1 -KD (infected with LV-sh DEC1 ). After 24 h of transfection or infection, the culture medium from each group was collected, centrifuged, and filtered to obtain conditioned medium (CM). Effect of CM from MC3T3-E1 cells with DEC1 -OE or KD on breast cancer cell migration by Transwell migration assays (D). The relative Cxcl12 mRNA levels in MC3T3-E1 cells (E). The CXCL12 amount in CMs from DEC1 -OE and KO in MC3T3-E1 cells by ELISA assays (F). G: Mechanism of DEC1 promoting breast cancer bone metastasis via transcriptional activation of CXCR4. Data are presented as mean ± standard deviation (all experiments were repeated at least three times). * P < 0.05, ** P < 0.01, *** P < 0.001, comparisons are shown in the figure. Data were analyzed using one-way ANOVA, and differences between groups were analyzed using Student's t -test. Abbreviation: EMT, epithelial-to-mesenchymal transition; KD, knockdown; OE, overexpression.

Article Snippet: CXCL12 enzyme-linked immunosorbent assay (ELISA) Kit (Cat. #KE10046) was from Proteintech (Wuhan, China).

Techniques: Expressing, Knock-Out, Immunohistochemical staining, Staining, Cell Culture, Plasmid Preparation, Infection, Transfection, Migration, Enzyme-linked Immunosorbent Assay, Activation Assay, Standard Deviation, Knockdown, Over Expression

Journal: International Journal of Immunopathology and Pharmacology

Article Title: Follow the time course of inflammation caused by intraperitoneal administration of multi-wall carbon nanotubes in mice

doi: 10.1177/03946320231176402

Figure Lengend Snippet: Cytokine level in the abdominal cavity lavage fluid. Two kinds of MWCNT (MWCNT-B and MWNT-7) were administered to abdominal cavity of the mice by intraperitoneal administration at a concentration of 1.0 mg (high dose) or 0.05 mg (low dose) per mouse. One milliliter of 0.5% Pluronic F-68 solution was also administered as control. The abdominal cavity lavage fluid was collected at 1 week, 1, 3 and 6 month(s) after intraperitoneal administration. Suspensions IL-6 and Cxcl-1 concentrations were determined using ELISA. The data are represented as mean ± SE. High dose of MWCNT-7 at 6 month was not tested (NT). * p < 0.05, ** p < 0.01 (vs. each time point of control mice, ANOVA and Dunnett’s test). # indicates under the analytical sensitivity (Cxcl-1: 6.5 pg/mL, IL-6: 4 pg/mL).

Article Snippet: The concentrations of Cxcl-1, Cxcl-2, mesothelin, and OPN in the serum and abdominal cavity lavage fluid were determined using a CXCL1 ELISA Kit (Proteintech Group, Inc., Rosemont, IL, USA), MIP-2/CXCL2 Mouse ELISA Kit (Thermo Fischer Scientific), mouse mesothelin ELISA kit (Abcam, Cambridge, UK), and mouse OPN (Osteopontin) ELISA Kit (Elabscience, Inc., Wuhan, Hubei, China), respectively.

Techniques: Concentration Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Immunopathology and Pharmacology

Article Title: Follow the time course of inflammation caused by intraperitoneal administration of multi-wall carbon nanotubes in mice

doi: 10.1177/03946320231176402

Figure Lengend Snippet: Osteopontin and mesothelin concentration in the abdominal cavity lavage fluid. Two kinds of MWCNT (MWCNT-B and MWNT-7) were administered to abdominal cavity of the mice by intraperitoneal administration at a concentration of 1.0 mg (high dose) or 0.05 mg (low dose) per mouse. One milliliter of 0.5% Pluronic F-68 solution was also administered as control. The abdominal cavity lavage fluid was collected at 1 week, 1, 3, and 6 month(s) after intraperitoneal administration of MWCNTs or 0.5% Pluronic F-68. Osteopontin and mesothelin concentrations were determined using ELISA. The data are represented as mean ± SE. High dose of MWCNT-7 at 6 month was not tested (NT). * p < 0.05, ** p < 0.01 (vs. each time point of control mice, ANOVA and Dunnett’s test). Means with different letters differ significantly at p < 0.05 (ANOVA and Newman–Keuls’s Test).

Article Snippet: The concentrations of Cxcl-1, Cxcl-2, mesothelin, and OPN in the serum and abdominal cavity lavage fluid were determined using a CXCL1 ELISA Kit (Proteintech Group, Inc., Rosemont, IL, USA), MIP-2/CXCL2 Mouse ELISA Kit (Thermo Fischer Scientific), mouse mesothelin ELISA kit (Abcam, Cambridge, UK), and mouse OPN (Osteopontin) ELISA Kit (Elabscience, Inc., Wuhan, Hubei, China), respectively.

Techniques: Concentration Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Immunopathology and Pharmacology

Article Title: Follow the time course of inflammation caused by intraperitoneal administration of multi-wall carbon nanotubes in mice

doi: 10.1177/03946320231176402

Figure Lengend Snippet: Aminotransferase (AST) concentration in the abdominal cavity lavage fluid. Two kinds of MWCNT (MWCNT-B and MWNT-7) were administered to abdominal cavity of the mice by intraperitoneal administration at a concentration of 1.0 mg (high dose) or 0.05 mg (low dose) per mouse. 1 milliliter of 0.5% Pluronic F-68 solution was also administered as control. The abdominal cavity lavage fluid was collected at 1 week, 1, 3 and 6 month(s) after intraperitoneal administration of MWCNTs or 0.5% Pluronic F-68. AST concentration was determined using ELISA. The data are represented as mean ± SE. High dose of MWCNT-7 at 6 months was not tested (NT). * p < 0.05, ** p < 0.01 (vs. each time point of control mice, ANOVA and Dunnett’s test).

Article Snippet: The concentrations of Cxcl-1, Cxcl-2, mesothelin, and OPN in the serum and abdominal cavity lavage fluid were determined using a CXCL1 ELISA Kit (Proteintech Group, Inc., Rosemont, IL, USA), MIP-2/CXCL2 Mouse ELISA Kit (Thermo Fischer Scientific), mouse mesothelin ELISA kit (Abcam, Cambridge, UK), and mouse OPN (Osteopontin) ELISA Kit (Elabscience, Inc., Wuhan, Hubei, China), respectively.

Techniques: Concentration Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Immunopathology and Pharmacology

Article Title: Follow the time course of inflammation caused by intraperitoneal administration of multi-wall carbon nanotubes in mice

doi: 10.1177/03946320231176402

Figure Lengend Snippet: Cytokine level in the abdominal cavity lavage fluid in early phase. Two kinds of MWCNT (MWCNT-B and MWNT-7) were administered to abdominal cavity of the mice by intraperitoneal administration at a concentration of 1.0 mg (high dose) per mouse. One milliliter of 0.5% Pluronic F-68 solution was also administered as control. The abdominal cavity lavage fluid was collected at 1, 3 days and 1 week after intraperitoneal administration. IL-6, Cxcl2, and TNF-α concentrations were determined using ELISA. The data are represented as mean ± SE * p < 0.05, ** p < 0.01 (vs. each time point of control mice, ANOVA and Dunnett’s test). # indicates under the sensitivity (Cxcl-2: 10 pg/mL).

Article Snippet: The concentrations of Cxcl-1, Cxcl-2, mesothelin, and OPN in the serum and abdominal cavity lavage fluid were determined using a CXCL1 ELISA Kit (Proteintech Group, Inc., Rosemont, IL, USA), MIP-2/CXCL2 Mouse ELISA Kit (Thermo Fischer Scientific), mouse mesothelin ELISA kit (Abcam, Cambridge, UK), and mouse OPN (Osteopontin) ELISA Kit (Elabscience, Inc., Wuhan, Hubei, China), respectively.

Techniques: Concentration Assay, Control, Enzyme-linked Immunosorbent Assay