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: Cell Regeneration

Article Title: Integrative analysis and experimental validation identify the role of CD44 and Nucleolin in regulating gliogenesis following spinal cord injury

doi: 10.1186/s13619-025-00253-x

Figure Lengend Snippet: Dynamic cell portion changes and intercellular communication analysis revealed SPP1 signaling pathway was critical in microglia after SCI. A , B Stacked bar plots depicting changes in the relative abundance of major cell types in spinal cord ( A ) and peripheral immune cell populations ( B ) across various time points. Astrocytes, microglia, OPCs, and MDMs show marked shifts, particularly in the acute (1~3 dpi) and subacute phases of SCI, the absence of 42 dpi stems directly from the source data ( GSE172167 ), where immune cell clusters were not identified or annotated at this specific time point in the original study. C Intercellular communication networks illustrate increased signaling complexity at 1 and 3 dpi compared to the sham condition. D Quantitative result of the total number of interactions in sham, 1 dpi, and 3 dpi samples, showing a significant increase in cell–cell interactions post-injury. E Heatmaps displaying the changes in signaling patterns for key cell types. SPP1 became prominent at 1 dpi. F Information flow of microglia indicated the SPP1 signal was significant

Article Snippet: After adhesion, recombinant mouse SPP1 protein (MCE, Cat No. HY- P71786 ) and recombinant mouse PTN protein (MCE, Cat No. HY- P71213 ) were separately administered to the microglia and astrocytes at concentrations of 0, 0.1, 0.5, and 1 μg/mL for a duration of 24 h. Subsequent to the stimulation period, the culture medium was carefully removed, and the cells were gently washed twice with PBS.

Techniques:

Journal: Cell Regeneration

Article Title: Integrative analysis and experimental validation identify the role of CD44 and Nucleolin in regulating gliogenesis following spinal cord injury

doi: 10.1186/s13619-025-00253-x

Figure Lengend Snippet: SPP1-CD44 signaling promotes microglial activation and inflammatory response. A SPP1 signaling pathway network showing interactions between microglia and other cell types. B A circular plot illustrating the interaction network of microglia with other cells in various ligand -receptor pairs, including Spp1 - Cd44 . C , D Violin plots showing expression levels of Spp1 and Cd44 across different cell types in sham (blue) and 1 dpi (red). Both genes show elevated expression in microglia following injury. E Violin plot of Cd44 expression in microglia subclusters, showing the upregulation in the wound healing and inflammatory response2 cluster at 1 dpi. F The microglia were sorted by flow cytometry and ( G ) Cd44 gene expression was detected by qRCR. H Flow cytometry analysis of CD44 positive microglia after SCI, showing a marked increase in CD44 + microglia during 7 dpi. I Immunofluorescence images of spinal cord lesion site stained for Iba1, CD44, SPP1, and merged with DAPI. White arrows indicate the co-stained CD44 + and SPP1 + signals in Iba1 positive microglia ( J ) Quantification results of CD44 + and SPP1 + in Iba1 positive microglia cells. K Using PLA to detect specific SPP1-CD44 interactions of spinal cord lesion site in situ. L Quantification of PLA results, the PLA signal is quantified and plotted as the area of PLA signal per Iba1 positive cell. M , N qRT-PCR showing dose- and time-dependent increases of Cd44 expression in BV2 microglia after recombinant SPP1 stimulation. O Representative images of PLA assay specific SPP1-CD44 interactions of BV2 microglia cells in vitro. P PLA signal was quantified and plotted as the area of PLA signal per cell. Q ELISA quantification of IL-6 levels in cell supernatant after SPP1 stimulation. R Western blot showing the time course of CD44 and p-NF-κB p65 protein expression in BV2 cells after SPP1 treatment. ( S – T ) Quantification of CD44 and p-NF-κB p65 protein levels. Data are presented as mean ± SEM. ( n = 3, * P < 0.05, ** P < 0.01, *** P < 0.001)

Article Snippet: After adhesion, recombinant mouse SPP1 protein (MCE, Cat No. HY- P71786 ) and recombinant mouse PTN protein (MCE, Cat No. HY- P71213 ) were separately administered to the microglia and astrocytes at concentrations of 0, 0.1, 0.5, and 1 μg/mL for a duration of 24 h. Subsequent to the stimulation period, the culture medium was carefully removed, and the cells were gently washed twice with PBS.

Techniques: Activation Assay, Expressing, Flow Cytometry, Gene Expression, Immunofluorescence, Staining, In Situ, Quantitative RT-PCR, Recombinant, In Vitro, Enzyme-linked Immunosorbent Assay, Western Blot

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

Journal: Frontiers in Immunology

Article Title: Osteopontin regulates right ventricular failure through integrin ανβ3/PERK/CHOP-dependent inflammatory and apoptotic pathways

doi: 10.3389/fimmu.2025.1569210

Figure Lengend Snippet: Clinical significance of serum OPN levels in right ventricular failure patients. (A) Serum OPN concentrations measured by ELISA in patients with normal right ventricular function (NF-RV, n=10) and pulmonary arterial hypertension-associated right heart failure (HF-RV, n=10). (B) Pearson correlation analysis between serum OPN and NT-proBNP levels in the HF-RV group (n=8). Solid line: linear regression fit; dashed lines: 95% confidence intervals. (C) Receiver Operating Characteristic (ROC) curve evaluating the diagnostic performance of serum OPN for right ventricular dysfunction. OPN, Osteopontin; NF-RV, Normal Right Ventricular Function; HF-RV, Right Heart Failure; PAH, Pulmonary Arterial Hypertension; NT-proBNP, N-terminal pro-Brain Natriuretic Peptide; ROC, Receiver Operating Characteristic; AUC, Area Under the Curve. Data presentation: Mean ± SD. Statistical significance: ** P < 0.01.

Article Snippet: Serum OPN levels were measured using the Human OPN ELISA Kit (EK0482, BOSTER) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Diagnostic Assay

Journal: Cells

Article Title: Involvement of Cyclooxygenase-2 in Establishing an Immunosuppressive Microenvironment in Tumorspheres Derived from TMZ-Resistant Glioblastoma Cell Lines and Primary Cultures

doi: 10.3390/cells13030258

Figure Lengend Snippet: List of primary antibodies used in the present study.

Article Snippet: rabbit monoclonal anti-osteopontin , 1:1000 , Boster Biological Technology, Pleasanton, CA, USA.

Techniques: