mural cells against pdgfr β  (R&D Systems)

Journal: bioRxiv

Article Title: Reduced Folate Carrier 1 (RFC1/Slc19a1) Suppression Exacerbates Blood-Brain Barrier Breakdown in Experimental Ischemic Stroke in Adult Mice

doi: 10.1101/2024.10.28.620539

Figure Lengend Snippet: A) PFA fixated 20 µm-thick brain sections of naive Swiss Albino mice was labelled with anti-RFC1 antibody (red), and α-SMA (green). RFC1 is expressed in α-SMA positive precapillary arterioles (arrows) as well as capillaries (arrowheads). B) RFC1 is not expressed in AQP4 positive astrocytic perivascular endfeet rather the RFC1 immunoreactivity is observed in spatially distinct regions. C) Isolated brain microvessel (< 9 μm diameter) preparations were immunohistochemically labelled with anti-RFC1 antibody (red), and colocalized with the vessel marker Lectin (green). The bump-on-a log pericyte shape can be observed as RFC1 positive (yellow arrows). D) The common pericyte marker PDGFR-β (green) colocalized with RFC1. Yellow arrow depicts a pericyte body appearing as a bump-on-a log on the microvessel. E) CD13 (green) also colocalized with RFC1. Nuclei were labeled with Hoechst 33258 (blue). Scale bars: 10 μm. F) RFC1 labeling in the post-mortem human cortex also showed that RFC1 is expressed in the endothelial cells and pericytes (yellow arrow) ( www.proteinatlas.org ). Scale bars: 20 μm

Article Snippet: Next, they were incubated overnight at + 4°C with primary antibodies against RFC1 (SLC19A1, MyBioSource MBS9134642 and Sigma-Aldrich AV44167; both produced in rabbit); for mural cells against PDGFR-β (R&D Systems, AF1042), CD13 (Acris Antibodies, AM26636AF-N); for endothelium CD31 (BD Bioscience, 550274).

Techniques: Isolation, Marker, Labeling

Journal: bioRxiv

Article Title: Prg4+ fibro-adipogenic progenitors in muscle are crucial for bone fracture repair

doi: 10.1101/2025.05.14.654160

Figure Lengend Snippet: (A) A representative fluorescence image of a Prg4ER/Td hindlimb. Mice at 2 months of age received Tam injections for 5 days and their hindlimbs were harvested at day 7. Top panel: a low magnification image containing tibiae (Tib), femur (fem), and joint tissue. Scale bar: 500 μm. Bottom panel: squares in the top panel are magnified to show joint (i, scale bar: 500 μm), muscle (Mus, ii, scale bar: 50 μm), cortical bone (Cb, iii, scale bar: 50 μm), and bone marrow (BM, iv, scale bar: 50 μm). M: meniscus; S: synovium; AC: articular cartilage; Ps: periosteum. (B) Fluorescent image of TA muscle stained for Prg4 mRNA by RNA FISH. Scale bar: 20 μm. (C) The Uniform Manifold Approximation and Projection (UMAP) plot of non-myogenic mesenchymal cells in mouse muscle. (D) Violin plot of Prg4 expression in muscle non-myogenic mesenchymal cell clusters. (E) Fluorescent images of TA muscle co-stained for Pdgfrα and WGA or Sca1 and WGA. Arrows point to Td+ Pdgfrα+ cells and Td+ Sca1+ cells. Scale bar: 20 μm. (F) Flow cytometer analysis of Td+ cells in muscle FAPs and non-FAPs. (G) Quantification of flow data. ***: p<0.001 vs Non-FAP, n=5 mice/time point. (H) Flow cytometer analysis of Td+ FAPs and Td-FAPs. (I) Quantification of flow data. ***: p<0.001 vs Td-, n=5 mice/time point.

Article Snippet: For immunofluorescence staining, sections were blocked by 3% bovine serum albumin (BSA) for 30_Jmin and incubated with rat anti-Sca1 (BioLegend, 122501), mouse anti-Pdgfrα (Santa Cruz, SC-398206), or rabbit anti-Perilipin (Cell Signaling, 9349) antibodies at 4°C overnight, followed by secondary antibody incubation for 1_Jh.

Techniques: Fluorescence, Staining, Expressing, Flow Cytometry