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 pharmacology

Article Title: Evaluation of the beneficial effects of a GABA-based product containing Melissa officinalis on post-inflammatory irritable bowel syndrome: a preclinical study.

doi: 10.3389/fphar.2024.1466824

Figure Lengend Snippet: FIGURE 11 Calprotectin levels in the preventive protocol of treatment (A) and curative protocol of treatment (B) in feces from control or DNBS rats treated with vehicle or GABA-Mo. Each column represents the mean ± SEM from eight animals.

Article Snippet: Fecal calprotectin levels were determined using a rat calprotectin ELISAkit (Cusabio EQ013485RA).

Techniques: Control

Journal: International immunopharmacology

Article Title: Oxidative stress induces ferroptosis in tendon stem cells by regulating mitophagy through cGAS-STING pathway.

doi: 10.1016/j.intimp.2024.112652

Figure Lengend Snippet: Fig. 1. Ferroptosis is involved in the oxidative damage of TSCs induced by H2O2. (A-B) Through Western blot and RT-PCR, the levels of GPX4, SLC7A11 protein and mRNA were assessed with or without the ferroptosis inhibitor Fer-1 (10 μM). (C) Detection of mitochondrial ultrastructure by TEM. (D) TSC cells were stained with Prussian blue. The levels of (E) Iron, (F) GSH, (G) MDA in TSC cells was examined using the appropriate test kits. By using ELISA assays, the level of (H) 4-HNE and (I) LPO were determined. (J) The ROS intensity in TSC cells was evaluated using ROS probe. (K) Western blot detected the protein levels of tenomodulin (n = 3); ***p < 0.001 and ###p < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: To evaluate intracellular 4-HNE, the ELISA kit (CUSA BIO; # CSBEQ027232RA) was utilized.

Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction, Staining, Enzyme-linked Immunosorbent Assay

Journal: International immunopharmacology

Article Title: Oxidative stress induces ferroptosis in tendon stem cells by regulating mitophagy through cGAS-STING pathway.

doi: 10.1016/j.intimp.2024.112652

Figure Lengend Snippet: Fig. 3. Inhibition of mitophagy attenuates ferroptosis induced by H2O2 in TSCs. TSCs cells were pretreated with Mdivi-1 (1 μM) for 2 h and then subjected to the prescribed concentration of H2O2 (100 μM) for a period of 24 h. (A) MTT experiment were used to test cell viability. (B) Using the relevant kits, the levels of GSH in TSC cells were measured. (C–D) To measure the level of intracellular iron, Prussian blue staining and an Iron Colorimetric Assay Kit were utilized. (E) MDA levels in TSC cells were determined using the relevant kits. (F-G) To measure the amount of 4-HNE and LPO in TSC cells, ELISA assays were employed. (H) The ROS con centration in TSC cells was measured using a ROS probe. (I) Western blot detected the protein levels of Tenomodulin (n = 3); **p < 0.01, ***p < 0.001, #p < 0.05, ##p < 0.01, and ###p < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: To evaluate intracellular 4-HNE, the ELISA kit (CUSA BIO; # CSBEQ027232RA) was utilized.

Techniques: Inhibition, Concentration Assay, Staining, Colorimetric Assay, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: International immunopharmacology

Article Title: Oxidative stress induces ferroptosis in tendon stem cells by regulating mitophagy through cGAS-STING pathway.

doi: 10.1016/j.intimp.2024.112652

Figure Lengend Snippet: Fig. 6. Interfering with cGAS attenuates ferroptosis induced by H2O2 in TSCs. TSC cells were treated with H2O2 for 24 h after receiving cGAS siRNA transfection for 12 h. (A) MTT experiment were used to test cell viability. (B) By using appropriate kits, the levels of GSH in TSC cells were determined. (C-D) To measure the level of intracellular iron, Prussian blue staining and an Iron Colorimetric Assay Kit were utilized. (E) To measure the amount of ROS present in TSC cells, a ROS probe was employed. (F) MDA levels in TSC cells were determined using the relevant kits. (G-H) To measure the amount of 4-HNE and LPO in TSC cells, ELISA assays were employed. (I) Western blot detected the protein levels of Tenomodulin. (n = 3); *p < 0.05, **p < 0.01, ***p < 0.001, #p < 0.05, ##p < 0.01, ###p < 0.001, ####p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: To evaluate intracellular 4-HNE, the ELISA kit (CUSA BIO; # CSBEQ027232RA) was utilized.

Techniques: Transfection, Staining, Colorimetric Assay, Enzyme-linked Immunosorbent Assay, Western Blot