Journal: Journal of Inflammation (London, England)

Article Title: The injury effect of osteopontin in sepsis-associated lung injury

doi: 10.1186/s12950-025-00430-4

Figure Lengend Snippet: rmOPN was supplemented after OPN silencing, and the protective effects in MH-s cells treated with LPS were reversed. ( A ) ELISAs were used to detected the concentration of IL-6 and TNF-α in cell supernatant of MH-s cells. ( B ) The mRNA expression levels of NLRP3, GSDMD, caspase1, ASC, IL-1β and IL-18 in MH-s cells. ( C ) Western blotting analysis of the relative expression of the NLRP3, GSDMD, caspase1, ASC, IL-1β and IL-18 in MH-s cells. The data are presented as the means ± standard deviations (S.D.). “*” indicates a difference between groups. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Osteopontin, OPN; recombinant mouse OPN, rmOPN; interleukin, IL; tumour necrosis factor, TNF; NOD-, LRR- and pyrin domain-containing 3, NLRP3; Gasdermin D, GSDMD; apoptosis-associated speck-like protein containing a CARD, ASC; lipopolysaccharide, LPS; Mouse alveolar macrophage cells, MH-s; enzyme-linked immunosorbent assay, ELISA; Quantitative Real-time reverse transcriptase-polymerase chain reaction, qPCR

Article Snippet: The cells were then divided into two groups: the experimental group, which was treated with 200 ng/mL of recombinant mouse OPN (rmOPN) protein (MCE, Osteopontin, HY-P78358, USA) for 30 min, and the control group, which received no such treatment.

Techniques: Concentration Assay, Expressing, Western Blot, Recombinant, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Polymerase Chain Reaction

Journal: bioRxiv

Article Title: Overcoming myeloid-driven resistance to CAR T therapy by targeting SPP1

doi: 10.1101/2025.04.01.646202

Figure Lengend Snippet: a, Schematic illustration demonstrating experimental design combining anti-SPP1 antibody and mIL13Rα2 CAR T cell therapy against YUMM JAK1/KO (n=5 per arm) and MC38 JAK1/KO tumors (n=5 per arm). b, Tumor volumes comparing combination therapy to mIL13Rα2 CAR T cell therapy alone and anti-SPP1 antibody treatment alone in YUMM JAK1/KO and MC38 JAK1/KO tumors. ** p<0.01, **** p<0.0001. Data are mean ± SEM. c, Cellular deconvolution of bulk RNA-seq from YUMM JAK1/KO tumors at day 17 (72 hours post-mIL13Rα2 CAR T cell administration on day 14), comparing tumors treated with a combination of anti-SPP1 and mIL13Rα2 CAR T cells, mIL13Rα2 CAR T cells alone, anti-SPP1 antibody alone, and an untreated control group. d, Representative immunofluorescent staining of CD68, COL1A1, and SPP1 in KLuc orthotopic glioma tumors. Bars indicate 50 µm. e, Schematic illustration of experimental design of KLuc orthotopic glioma model treated with combination of anti-SPP1 antibody and mIL13Rα2 CAR T cell therapy compared to mIL13Rα2 CAR T cell therapy alone and anti-SPP1 antibody alone groups (n=5 per arm). f, Survival analysis revealed significantly longer survival of KLuc harboring mice treated with combination therapy compared to mIL13Rα2 CAR T alone, anti-SPP1 antibody alone treated groups. Data are mean ± SEM. g, Flux values revealed significantly lower bioluminescence values in KLuc harboring mice treated with combination therapy compared to mIL13Rα2 CAR T alone, anti-OPN antibody alone treated groups. h, Mice heads showing the flux values comparing mIL13Rα2 CAR T and anti-OPN antibody combined treated group to mIL13Rα2 CAR T alone group.

Article Snippet: The simultaneous treatment group was given an initial dose of anti-OPN antibody (bioXcell Clone: 100D3) on the day of CAR T cell therapy, followed by five more doses to ensure parity in antibody delivery across groups, all given intraperitoneally (IP).

Techniques: RNA Sequencing, Control, Staining

Journal: Nature Communications

Article Title: Single-cell and spatial transcriptome analyses reveal tumor heterogeneity and immune remodeling involved in pituitary neuroendocrine tumor progression

doi: 10.1038/s41467-025-60028-5

Figure Lengend Snippet: a CCK-8 cell viability assay of GH3, AtT-20, and TtT/GF cells treated with OPN or PBS ( n = 5 biological replicates). Data are presented as mean values +/- SD. b Representative images of the wound healing assay, and ( c ) quantitative analysis of TtT/GF cells treated with OPN or PBS ( n = 3 biological replicates). Data are presented as mean values +/- SD. d Transwell invasion assay and ( e ) quantitative analysis of GH3, AtT-20, and TtT/GF cells treated with OPN or PBS ( n = 3 biological replicates). Data are presented as mean values +/- SD. f Immunofluorescence images of PitNET markers (SYN, SF1, FSH) in PitNET organoids from two independent patients ( n = 3 biological replicates). g Transwell invasion assay and ( h ) quantitative analysis of primary pituitary tumor cells cultured from two independent patients, treated with OPN or PBS ( n = 3 biological replicates). Data are presented as mean values +/- SD. i Bright-field images illustrating representative phenotypes of three independently derived PitNET organoids from each of the two patients treated with OPN or PBS. Arrows indicate cell protrusions in PitNET organoids. j CellTiter-Glo luminescent cell viability assay of PitNET organoids treated with OPN or PBS ( n = 3 biological replicates). Data are presented as mean values +/- SD. k Immunofluorescence images of Ki67 in PitNET organoids from two patients treated with OPN or PBS ( n = 3 biological replicates). For all panels, scale bar, 100 µm. Two-sided unpaired t -test was performed for ( a − j ). Data are presented as mean values +/- SD for ( a − j ). Source data are provided as a Source Data file for ( a − j ).

Article Snippet: After overnight attachment, cells were treated with either PBS as a control or OPN (Rat OPN protein, MedChemExpress, HY- P79222 ; Recombinant mouse OPN protein, bioswamp, RPH01180 .) at a final concentration of 5 μg/ml.

Techniques: CCK-8 Assay, Viability Assay, Wound Healing Assay, Transwell Invasion Assay, Immunofluorescence, Cell Culture, Derivative Assay, Cell Viability Assay