Journal: Aging (Albany NY)

Article Title: Age dependent increase in the levels of osteopontin inhibits skeletal muscle regeneration

doi:

Figure Lengend Snippet: ( A ) Blood sera were collected from young and old mice, either non-injured or at 3 days and 5 days after muscle injury and OPN levels were quantified by ELISA. Resting young and old blood serum levels do not show significant age-specific difference, while both at 3 days and 5 days after muscle injury old mice have higher levels of serum osteopontin than young (n=3-6, ± SEM, p**≤0.05). ( B ) Tibialis Anterior (TA) muscle sections from uninjured (resting), 3DPI and 5DPI muscle were co-immunostained for Laminin (to visualize the myofiber periphery) and OPN. OPN is undetectable in the uninjured skeletal muscle or young and old mice. In contrast, at 3 days post injury both young and old muscles show pronounced OPN at the site of injury and old muscle has considerably more OPN than young. By 5 days post injury, OPN becomes greatly diminished at the site of injury / regeneration in young muscle, but old muscle with its poor repair displays sustained OPN presence. At 3 days post injury myofibers ( C ) and myogenic stem cells ( D ) obtained from young and old mice were analyzed for OPN expression by western blotting and qRT-PCR ( F ) Old myogenic stem cells expressed higher levels of OPN as compared to young both at protein and mRNA levels, while the levels of OPN did not change with age in myofibers. (quantified in E; n=3, ± SEM, p**≤0.05).

Article Snippet: CD11b (rat monoclonal), goat normal IgG and osteopontin (goat polyclonal) for immunostaining and in vivo neutralization were from R&D, Pax7 and eMyHC was from DSHB; secondary HRP antibodies were purchased from Santa Cruz.

Techniques: Enzyme-linked Immunosorbent Assay, Muscles, Expressing, Western Blot, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Single-cell RNA sequencing identifies phenotypically, functionally, and anatomically distinct stromal niche populations in human bone marrow

doi: 10.1101/2022.01.26.477664

Figure Lengend Snippet: (A-D, F-G) UMAP (as in ) illustration of the normalized expression of selected ligand and receptor gene pairs. Red dashed lines in figures A, B and D mark the erythroid clusters. The blow-up shown in (G) is to better visualize PDGFA- and PDGFB-expressing cells. (E) Formalin-fixed, paraffin-embedded (FFPE) human BM slides were sequentially stained for DAPI (blue), CD45 (yellow), SPP1 (red), CD271 (pink), and NCAM1 (cyan) and scanned with an OlympusVS120 slide scanner. Lower panel: Single staining for each marker is shown as indicated under each image. Scale bars represent 50 µm. White lines indicate bone lining regions. b, Bone.

Article Snippet: For SPP1 staining, after deparaffinization, rehydration, and antigen retrieval, slides were permeabilized with 0.1% Triton X100 (Sigma Ultra, T9284) solution at 37°C for 15 minutes, blocked using donkey serum (Jackson Immuno Research, 017-000-121), and stained overnight at 4°C with goat anti-human SPP1 antibody (R&D Systems, AF1433-SP), followed by secondary staining with donkey anti-goat AF594 (Jackson ImmunoResearch, 706-585-147) and staining with CD56-AF647 plus DAPI.

Techniques: Expressing, Formalin-fixed Paraffin-Embedded, Staining, Marker

Journal:

Article Title: Inhibition of p53-induced apoptosis without affecting expression of p53-regulated genes

doi: 10.1073/pnas.1031695100

Figure Lengend Snippet: Verification of IL-6-induced expression of Bcl3 and Spp1. (a) Real-time quantitative PCR result of expression of Bcl3 by p53 or p53+IL-6. (b) Western blot analysis of Spp1 (osteopontin) in M1-t-p53 cells cultured at 32°C(Upper) or 37°C (Lower) without (-) or with (+) IL-6 or TG.

Article Snippet: The blots were incubated with goat anti-mouse osteopontin (Spp1) antibody (1:200) followed by horseradish peroxidase-conjugated rabbit anti-goat IgG antibody (1:2,000) (both from Santa Cruz Biotechnology).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Cell Culture

Journal:

Article Title: Inhibition of p53-induced apoptosis without affecting expression of p53-regulated genes

doi: 10.1073/pnas.1031695100

Figure Lengend Snippet: Regulation of some antiapoptotic, proapoptotic, and differentiation-associated gene expression by IL-6 and TG

Article Snippet: The blots were incubated with goat anti-mouse osteopontin (Spp1) antibody (1:200) followed by horseradish peroxidase-conjugated rabbit anti-goat IgG antibody (1:2,000) (both from Santa Cruz Biotechnology).

Techniques: Expressing

Journal: bioRxiv

Article Title: Fluid-Niche and Microglial Signatures Prime Robust Intraventricular Macrophage Response to Blood During Brain Development

doi: 10.1101/2025.10.02.679906

Figure Lengend Snippet: ( A ) Myeloid cell population clustering from E16.5 ChP. ( B ) CellChat visualization of inferred outgoing signals from epithelial, mesenchymal, and endothelial cells to ChP macrophages. ( C ) Incoming vs. outgoing signal strength across populations. ( D ) Heatmap of selected differentially expressed genes from bulk RNA sequencing of sorted P7 stromal vs epiplexus macrophages. ( E ) Representative image of an E14.5 brain sections stained with SPP1. Scale bar = 10 µm. ( F ) UMAP of other myeloid population signatures reflected in the various ChP macrophage populations. ( G ) Schematic of choroid plaque development from E12.5-E16.5. ( H ) Representative image of an E12.5 brain section. Scale bar = 200 µm. Inset: Magnified view of choroid plaque. Scale bar = 50 µm. ( I ) Representative image of an E17.5 and E18.5 brain section showing the residual choroid plaque and underlying macrophages. Scale bar = 250 µm; 25 µm for inset. ( J ) Representative image of a P0 brain section showing the roughly equivalent region following the crossing of the corpus callosum as well as the lateral ventricle and ChP. Scale bar = 250 µm; 25 µm for inset.

Article Snippet: 488 (ThermoFisher, 53-0443-82, 1:100), Goat anti-OPN (SPP1) (Novus Biologicals, AF808, 1:100), Goat anti-GPNMB (R&D Systems, AF2330, 1:500), Rat anti-LY76 (Ter119)(Abcam, ab91113, 1:500), Chicken anti-IBA1 (Synaptic Systems, 234-009, 1:500), Rabbit anti-AQP1 (Millipore Sigma, AB2219, 1:100), Rabbit anti-MRC1 (CD206) (Abcam, ab64693, 1:250), Rabbit anti-HMOX1 (Proteintech, 10701-1-AP, 1:250).

Techniques: RNA Sequencing, Staining