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: Kidney international

Article Title: Vitamin D receptor agonists increase klotho and osteopontin while decreasing aortic calcification in mice with chronic kidney disease fed a high phosphate diet

doi: 10.1038/ki.2012.322

Figure Lengend Snippet: (A) Arterial medial osteopontin (OPN) levels were increased by VDRA treatment. (i) OPN expression was low but detectable in the aortic media of CKD mice fed a high phosphate diet (CKD+HP). VDRA treatment increased smooth muscle cell expression of OPN in the aortic media in (ii) CKD+HP+C30 animal; and (iii) CKD+HP+P300 animal. Arrows point to aortic medial cells expressing OPN. Scale bars are 30 μm and objective is 40X. (B) Quantitation of OPN immunostaining showed no OPN expression in aortas from non-CKD controls, weak staining in CKD high phosphate-fed mice, and increased levels in VDRA-treated CKD mice (mean ± s.e.m., n =3 for all except CKD+HP+paricalcitol, n =5 where P100 and P300 samples were grouped). (C) Treatment of cultured VSMCs with 50 nM paricalcitol increased OPN levels in the media (significantly higher levels by ELISA at 48hr compared to time zero, ** P <0.001). Klotho (2 ng/mL) with/without FGF23 (2 ng/mL) did not upregulate OPN secretion. Three wells were sampled per time-point per treatment group, data are mean ± s.e.m. EtOH = ethanol control, KL = klotho, P50 = paricalcitol 50 nM.

Article Snippet: The following assays were used: QuantiChromTM Urea Assay Kit (BioAssay Systems, Hayward, CA) for BUN; o -cresolphthalein complexone kit from Teco Diagnostics (Anaheim, CA) for calcium; standard bioanalyzer at Phoenix Central Laboratory (Everett, WA) for phosphate; mouse FGF23 C-terminus ELISA kit (Immutopics, San Clemente, CA); DuoSet mouse osteopontin ELISA kit (R&D Systems, Minneapolis, MN); mouse intact PTH enzyme-linked immunosorbent assay kits (ALPCO Diagnostics, Salem, NH and Immutopics).

Techniques: Expressing, Quantitation Assay, Immunostaining, Staining, Cell Culture, Enzyme-linked Immunosorbent Assay, Control