Journal: Signal Transduction and Targeted Therapy

Article Title: Single-cell transcriptomic analysis reveals gut microbiota-immunotherapy synergy through modulating tumor microenvironment

doi: 10.1038/s41392-025-02226-7

Figure Lengend Snippet: Synergistic effects of gut microbiota and ICI treatment on SPP1. a Violin plots reveal Spp1 expression levels in myeloid subtypes across four groups. b Forest plot reveals the association between SPP1 gene expression levels and the prognosis of major cancer types in the TCGA dataset. c Kaplan–Meier survival curves show the correlation between SPP1 expression levels and prognosis in two immunotherapy cohorts. d Box plots indicate the levels of osteopontin measured by ELISA in the serum of mice across four groups. Each group consists of 4 mice. e Network diagram illustrates the differential interaction strength of the Spp1-related pathway between two cell types in the PW group compared to the others. f Bubble plot displays the interaction strength of the Spp1-related pathways between tumor-associated macrophages (TAMs)/NK cells and MC38 tumor cells in the PW group compared to the others. g Network diagram shows the differential interaction strength of the Spp1-related pathway between myeloid subtypes and MC38 cells in the PW group compared to the others. h Network diagram illustrates the differential interaction strength of the overall signaling between myeloid subtypes and MC38 cells in the PW group compared to the others

Article Snippet: The concentration of osteopontin (OPN) in mouse serum samples was quantified using the Mouse/Rat Osteopontin ELISA Kit (KE10046; Proteintech) with the following procedure.

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

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