Journal: iScience

Article Title: In vitro vascularized immunocompetent patient-derived model to test cancer therapies

doi: 10.1016/j.isci.2023.108094

Figure Lengend Snippet:

Article Snippet: Mouse anti-CD31 , Cell Applications , Cat# CB13678.

Techniques: Plasmid Preparation, Recombinant, Lysis, RNA Extraction, Amplification, DC Protein Assay, Enzyme-linked Immunosorbent Assay, Sequencing, Software, Fluorescence, Microscopy

Journal: Communications Biology

Article Title: Angular difference in human coronary artery governs endothelial cell structure and function

doi: 10.1038/s42003-022-04014-3

Figure Lengend Snippet: FE-SEM (top) based structural and surface level analysis of EC exposed to D-Flow using 30°, 60° and 80° microchannels. Magnification: ×5000, Scale: 10 µm. Flow exposed (4 h; 30°, 60° and 80° microchannels) EA.hy926 cells stained for cell-cell contact markers CD31 (second) and CD144 (penultimate). DAPI staining is shown in blue. Circularity of such cells was assessed using ImageJ. Cell circularity AU values per individual cells are indicated together with mean. Total number of cells, n ≥ 30. Values represent the mean ± SD. * p < 0.05, ** p < 0.01 and *** p < 0.001, by one-way ANOVA. Magnification: ×50/×63, Scale: 80 µm/20 µm. Cell cytoskeletal F-actin (bottom) staining for EA.hy926 exposed to D-Flow (D-variable or D-fixed regions) in 30°, 60° and 80° microchannels, using phalloidin. DAPI staining is shown in blue. Magnification: ×63, Scale: 20 µm.

Article Snippet: Following a 10 min fixation with 4% paraformaldehyde (PFA) and permeabilization using 0.1% Triton X-100 (5 min), the glass slides were blocked with 2% bovine serum albumin for 1 h; and further incubated overnight at 4 °C with eNOS Rabbit mAb (1:100, #32027), ICAM1 Rabbit pAb (1:500; #4915), CD31 (PECAM-1) Mouse mAb (1:1000; #3528) (Cell Signaling Technology, Danvers, USA), and CD144 (VE-Cadherin) Mouse mAb (1:500; #SC-9989; Santa Cruz Biotechnology, Dallas, USA) primary antibodies.

Techniques: Staining

Journal: Materials Today Bio

Article Title: Delivered baicalein immunomodulatory hydrogel with dual properties of pH-responsive and anti-infection orchestrates pro-regenerative response of macrophages for enhanced hypertrophic scars therapy

doi: 10.1016/j.mtbio.2025.102160

Figure Lengend Snippet: Functional performance and biocompatibility evaluation of hydrogels. (A) Photographs of MRSA and E.coil colonies, and (B, C) corresponding quantitative calculation of antibacterial rates after treatments of different hydrogels. (D) Representative scratch assay images of L929 cells treated with different hydrogels and (E) corresponding quantitative analysis of cell migration rates based on scratch area. ELISA was performed to assess the expression of CD31 (F) and VEGF (G) in HUVEC cells following treatment with various hydrogels. (H) Photographs of hemolysis assays and (I) statistically analyzed hemolysis rates. (J) Cytocompatibility of hydrogels and in vivo hematological parameters, including (K)white blood cell (WBC) count -, (L)red blood cell (RBC) count , and (M)platelet count. n = 3, ∗P < 0.05, ∗∗P < 0.01 and ∗∗∗P < 0.001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Following seeding in 12-well culture plates, HUVECs were maintained for 72 h in experimental media supplemented with different test materials, after which the supernatants were collected, centrifuged, and analyzed for CD31 (E-EL-H6227, Elabscience) and VEGF (E-EL-H0111, Elabscience) concentrations.

Techniques: Functional Assay, Wound Healing Assay, Migration, Enzyme-linked Immunosorbent Assay, Expressing, In Vivo

Journal: Nature Aging

Article Title: Anti-uPAR CAR T cells reverse and prevent aging-associated defects in intestinal regeneration and fitness

doi: 10.1038/s43587-025-01022-w

Figure Lengend Snippet: a , Surface uPAR expression as determined by flow cytometry on isolated intestinal crypts from young (3 months) and old (20 months) mice ( n = 3 per group). b , Percentage of uPAR + cells that are either EpCAM + CD45 − or EpCAM − CD45 + as determined by flow cytometry on isolated intestinal crypts from old (20 months) mice ( n = 3 per group). c , Surface uPAR expression in SPiDER-β-gal + cells as determined by flow cytometry on isolated intestinal crypts from old (20 months) mice ( n = 3 per group). d , Representative co-immunofluorescence of uPAR (red) and EdU (green) in the proximal jejunum of aged (20 months old mice) ( n = 3 mice). White arrows signal uPAR + EdU − cells. e , Percentage of uPAR + cells that are EdU + or EdU − in the proximal jejunum of aged (20 months) mice ( n = 3 mice). f , Representative co-immunofluorescence of uPAR (red) and p21 (green) in the proximal jejunum of aged (20 months) mice ( n = 3 mice). White arrows signal uPAR + cells. g , Percentage of uPAR + cells that are p21 + or p21 − in the proximal jejunum of aged (20 months) mice ( n = 3 mice). h – k , uPAR + and uPAR − cells from isolated intestinal crypts of duodenum, jejunum and ileum (whole small intestine) from old (20 months) mice were FACS sorted and subjected to scRNA-seq ( n = 4 mice per group pooled into two replicates per group). i , Uniform Manifold Approximation and Projection (UMAP) visualization of small intestinal cell types generated by 10X chromium protocol. Color scale indicates differences in density of cellular populations between uPAR + and uPAR − cells. j , Pathway analysis using enrichR comparing differentially expressed genes between uPAR + versus uPAR − cells in scRNA-seq data. Size scale represents number of genes in each ontology, and color scale represents degree of significance. k , UMAP visualization of small intestinal cell types generated by 10X chromium protocol. Color scale indicates log 2 fold change (log2FC) in senescence signature between uPAR + and uPAR − cells. Right: quantification of the proportion of uPAR + and uPAR − cells contributing to the senescence signature. l – n , scRNA-seq of small intestinal non-immune cell types in the whole small intestine of young (25–30 years old) and old (65–70 years old) subjects generated by 10X chromium protocol ( n = 1 per group). m , Split-violin plot indicates the expression level of PLAUR in the ISC and epithelial lineage. Boxplots display median (center line) and interquartile range (box). n , Pathway analysis using enrichR comparing differentially expressed genes between non-immune PLAUR + vs PLAUR − cells in scRNA-seq data. Size scale represents number of genes in each ontology, and color scale represents degree of significance. o – q , scRNA-seq of small intestinal immune cell types in the whole small intestine of young (25–30 years old) and old (65–70 years old) subjects generated by 10X chromium protocol ( n = 1 per group). p , Split-violin plot indicates the expression level of PLAUR in B cells, myeloid cells and T cells. Boxplots display median (center line) and interquartile range (box). q , Pathway analysis using enrichR comparing differentially expressed genes between immune PLAUR + versus PLAUR − cells in scRNA-seq data. Size scale represents number of genes in each ontology, and color scale represents degree of significance. r , Multiplex immunofluorescence of uPAR, ki-67, p21, γH2A.X, cleaved caspase-3, CD45, CD31, E-cadherin and DAPI in human intestinal samples from subjects aged 51–91 years ( n = 3 subjects). Green arrows highlight uPAR + p21 + cells, pink arrows highlight uPAR + γH2A.X + cells. s , Percentage of cells in the tissues from t that are uPAR + CD45 + uPAR + , CD68 + uPAR + or E-Cadherin + uPAR + ( n = 3 subjects). t , Percentage of E-Cadherin + uPAR + from t that are Ki-67 − , p21 + , Ki-67 − and p21 + , γH2A.X + , or Ki-67 − , p21 + and γH2A.X + ( n = 3 subjects). Shown are results of one independent experiment ( a –t ). Data are mean ± standard error of the mean (s.e.m.) ( a –c , e , g , s–t ). Significance was determined using a two-tailed unpaired Student’s t -test ( a –c , e , g ), two-tailed Fischer’s exact test ( j , n , q ) or two-tailed Wilcoxon rank-sum test (* P < 0.05,** P < 0.01, *** P < 0.001, **** P < 0.0001 ( m , p ). Illustration was created with Biorender.com ( h ).

Article Snippet: The following antibodies were used: uPAR (AF807, R&D), AF555-Ki-67 (558617, BD Biosciences), AF647-γH2A.X (ab195189, Abcam), AF488-E-cadherin (3199S, Cell Signaling Technology), AF647-p21 (8587S, Cell Signaling Technology), AF488-CD31 (42777, Cell Signaling Technology), AF555-CD45 (19744, Cell Signaling Technology), AF750 cleaved caspase-3 (97774S, Cell Signaling Technology), and AF555-donkey anti goal ( A21432 , Invitrogen).

Techniques: Expressing, Flow Cytometry, Isolation, Immunofluorescence, Generated, Multiplex Assay, Two Tailed Test

Journal: Nature

Article Title: Capillary cell-type specialization in the alveolus.

doi: 10.1038/s41586-020-2822-7

Figure Lengend Snippet: Fig. 1 | Two stable, intermingled alveolar capillary cell types. a, Alveolar capillaries in adult mouse lung immunostained for PECAM1. b, t-distributed stochastic neighbour embedding (t-SNE) plot of endothelial cell populations annotated in scRNA-seq data for adult mouse lung13. c, Heat map of expression of capillary subset markers and the general endothelial marker Cldn5 in individual capillary cells. CPM, counts per million. d–f, Single-molecule fluorescent in situ hybridization (smFISH) for the capillary subset markers Apln (d, f) or Ednrb and Car4 (aCap) (e), and Aplnr (gCap) (d–f), in adult mouse lung. Images in d (right) and f show individual aCap and gCap cells. g, Relative abundance of aCap cells, gCap cells and cells that co-express aCap and gCap markers (intermediate (IM) cells) in lungs from 3-month-old (young) and 24-month-old (aged) mice (data shown as mean; n = 500 cells scored per mouse; 2 mice per group). h, i, Co-expression of tdTomato lineage label (asterisks) and aCap marker Ednrb but not gCap marker Aplnr (h), or gCap marker Aplnr but not aCap marker Apln (i), in lungs collected six months after mature aCap (h) or gCap (i) cells were lineage-labelled. Blue, DAPI. Scale bars, 10 μm.

Article Snippet: CD34 (BD Biosciences, 347660):https://www.bdbiosciences.com/eu/applications/research/clinical-research/oncology-research/ 3 nature research | reporting sum m ary O ctober 2018 blood-cell-disorders/surface-markers/human/purified-mouse-anti-human-cd34-my10/p/347660 Endomucin (Invitrogen, eBioV.7C7, 14-5851-82): https://www.thermofisher.com/antibody/product/Endomucin-Antibody-cloneeBioV-7C7-V-7C7-Monoclonal/14-5851-82 Integrin alpha8 (R&D, AF4076): https://www.rndsystems.com/products/mouse-rat-integrin-alpha8-antibody_af4076 Pecam1 (rat anti-mouse; BD Biosciences, 553370): https://www.bdbiosciences.com/ds/pm/tds/553370.pdf Pecam1 (mouse anti-human; R&D, BBA7): https://www.rndsystems.com/products/human-cd31-pecam-1-antibody-9g11_bba7 tdTomato (Rockland, 600-401-379): https://rockland-inc.com/store/Antibodies-to-GFP-and-Antibodies-to-RFP-600-401-379O4L_24299.aspx VE-Cadherin (R&D, AF938): https://www.rndsystems.com/products/human-ve-cadherin-antibody_af938 The following primary antibodies were used to stain turtle and/or alligator tissue.

Techniques: Expressing, Marker, In Situ Hybridization