recombinant opn protein  (MedChemExpress)

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: 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