Journal: Biology of reproduction

Article Title: CD9 is a surface marker on mouse and rat male germline stem cells.

doi: 10.1095/biolreprod.103.020867

Figure Lengend Snippet: FIG. 1. Expression of CD9 in mouse testis. A) Characterization of anti-CD9 antibody-purified mouse testis cells using flow cytometry. Unselected (upper row) and anti-CD9 antibody-selected (lower row) testis cells were stained with FITC-conjugated streptavidin, PE-conjugated anti-a6-integrin antibody, or APC-conjugated anti-c-kit antibody, and the fluorescence was compared with that of controls. The dotted lines show the range of fluores- cence in the controls. Note the increased percentages of all three antigens in the anti-CD9 antibody-selected cell population. B) Immunohistological staining of CD9 antigen in mouse testis. Positive cells were found on the basement membrane of seminiferous tubules (arrows). Staining was also observed in the interstitial cells. Bar 5 50 mm (section). Stain: DAB followed by hematoxylin.

Article Snippet: Mouse anti-rat CD9 antibody was used with goat anti-mouse IgG microbeads (25 ml; Miltenyi Biotec) to select cells expressing rat CD9 molecules.

Techniques: Expressing, Cytometry, Staining, Membrane

Journal: Biology of reproduction

Article Title: CD9 is a surface marker on mouse and rat male germline stem cells.

doi: 10.1095/biolreprod.103.020867

Figure Lengend Snippet: FIG. 2. Expression of CD9 on mouse spermatogonial stem cells. A) Gross and histological appearance of recipient testes after transplantation of LacZ-marked donor testis cells. Left: Testis from a W recipient mouse 2 mo after transplantation of anti-CD9 antibody-selected cells. Middle: Testis from a W recipient mouse 2 mo after transplantation of unselected testis cells. Right: Histological section from a W recipient testis 2 mo after transplantation of anti-CD9-antibody-selected testis cells. Note the normal appearance and organization of spermatogenesis. Stain: X-gal followed by hematoxylin and eosin. B) Enhanced colonization of recipient testis by anti-CD9 antibody-selected mouse testis cells. The degree of coloni- zation in three experiments is indicated by the number of individual blue colonies. The values (mean 6 SEM) for anti-CD9 antibody-selected cells and unselected cells were 5.5 6 0.8 (n 5 16) and 0.8 6 0.2 (n 5 18) cells per 3 3 104 injected cells, respectively. Bars 5 1 mm (testes) and 50 mm (section).

Article Snippet: Mouse anti-rat CD9 antibody was used with goat anti-mouse IgG microbeads (25 ml; Miltenyi Biotec) to select cells expressing rat CD9 molecules.

Techniques: Expressing, Transplantation Assay, Staining, Injection

Journal: Biology of reproduction

Article Title: CD9 is a surface marker on mouse and rat male germline stem cells.

doi: 10.1095/biolreprod.103.020867

Figure Lengend Snippet: FIG. 3. Expression of CD9 on rat spermatogonial stem cells. A) Characterization of anti-CD9 antibody-purified rat testis cells using flow cytometry. Unselected (left) and anti-CD9 antibody-selected (right) testis cells were stained with Cy5-conjugated anti-mouse IgG antibody, and the fluorescence was compared with similar cell controls. The dotted lines show the control range of fluorescence. Note the increase in percentage of CD9-positive cells after magnetic selection. B) The appearance of recipient testes after transplantation of green fluorescent protein-marked donor testis cells. Seminiferous tubules were dissected using fine forceps. Left: Testis from a busulfan-treated nude recipient mouse 3 mo after transplantation of unselected testis cells (8.2 3 104 cells injected). Right: Testis from a busulfan-treated nude recipient mouse 3 mo after transplantation of anti-CD9 antibody-selected cells (2.7 3 104 cells injected). Note the increase in number of colonies of CD9-positive cells despite the lower concentration of injected cells. Bar 5 100 mm. C) Enhanced colonization of recipient testis by anti-CD9 antibody-selected rat testis cells. The number of individual fluorescent colonies represents the degree of colonization in three experiments. The values (mean 6 SEM) for anti-CD9 antibody-selected and unselected cells were 15.8 6 4.2 (n 5 16) and 3.1 6 0.9 (n 5 13) cells per 105 injected, respectively.

Article Snippet: Mouse anti-rat CD9 antibody was used with goat anti-mouse IgG microbeads (25 ml; Miltenyi Biotec) to select cells expressing rat CD9 molecules.

Techniques: Expressing, Cytometry, Staining, Control, Selection, Transplantation Assay, Injection, Concentration Assay

Journal: mBio

Article Title: Candida albicans Induces Cross-Kingdom miRNA Trafficking in Human Monocytes To Promote Fungal Growth

doi: 10.1128/mbio.03563-21

Figure Lengend Snippet: EVs attach to the surface of C. albicans . (a) Human CD9-positive MEV Ca (left) or MEV (right) (each from 5 × 10 5 monocytes) adhered to the surface of C. albicans , as seen in fixed imaging (CLSM). Blue, C. albicans ; red, CD9. Bars, 10 μm. Data are representative of n = 4 independent experiments. (b) C. albicans -induced EVs bound more abundantly than EVs from untreated PBMCs to C. albicans . Stained C. albicans are shown to exclude staining effects. (c) C. albicans -induced EVs from PBMCs exposed CR1, CD9, and CD63 on the surface. (d) Inhibition of CR1 on the EVs substantially reduced the attachment of EVs to C. albicans . (e) EVs from C. albicans -induced PBMCs (PEV Ca ) are substantially reduced when C. albicans is not opsonized (-op). Data in panels b, d, and e represent mean values ± SD, unpaired two-tailed t test; n = 4 independent experiments.

Article Snippet: CD14 was stained with Alexa Fluor 488 anti-human CD14 antibody (no. 367130; BioLegend) (1:100), CD9 was stained with Alexa Fluor 647 anti-human CD9 antibody (no. MCA469A647T; Bio-Rad) (1:1,000), TLR4 was stained with mouse anti-TLR4 antibody (no. NBP1-51697; R&D Systems) (3 μg/ml) and Alexa Fluor 647 goat anti-mouse IgG (H+L) secondary antibody (no. A-21235; Thermo Fisher) (1:500), and CD11b was stained with rabbit anti-CD11b antibody (no. 133357; Abcam) and Alexa Fluor 488 goat anti-rabbit IgG (H+L) secondary antibody.

Techniques: Imaging, Staining, Inhibition, Two Tailed Test

Journal: mBio

Article Title: Candida albicans Induces Cross-Kingdom miRNA Trafficking in Human Monocytes To Promote Fungal Growth

doi: 10.1128/mbio.03563-21

Figure Lengend Snippet: hsa-miR-24-3p increases C. albicans growth. (a) Growth of C. albicans was measured after the fungus was incubated with MEV Ca or MEV. Growth was monitored over 30 h in a plate reader, which measured the OD every hour. Data are presented as mean values ± SD; n = 3 different donors. (b) C. albicans hyphae grew in the presence of MEV Ca but not MEV, as observed by CLSM. Blue, C. albicans . Bars, 10 μm. Data are representative of n = 4 independent experiments. (c) C. albicans hyphae, measured using the ZEN 2011 software, were significantly longer in the presence of MEV Ca than MEV. Data are presented as mean values ± SD, P < 0.0001; n = 17; from n = 3 different experiments. (d) RNA from CD9-positive vesicles also migrated into C. albicans , as observed in CLSM. (e) RNA from MEV Ca migrated into C. albicans , as observed by 3D SIM. For panels d and e, data are representative of n = 3 independent experiments. Blue, C. albicans ; green, RNA; red, CD9. Bars, 10 μm. (f) hsa-miR-24-3p together with fluorescently labeled hsa-miR-24-3p sensor migrates into C. albicans (lower) but significantly less when hsa-miR24-3p was inhibited in monocytes (upper). (g) Mean values ± SD of fluorescence from 5 independent experiments as measured by a plate reader are shown on the right ( P = 0.003, unpaired two-tailed t test). (h and i) Hyphal growth of C. albicans in the presence of hsa-miR-24-3p. Blue, C. albicans . Bars, 10 μm. Data are presented as mean values ± SD; P < 0.0001; n = 20; n = 4 different experiments. (j and k) Incubation with hsa-miR-24-3p increased growth of both C. albicans and human hepatic cancer cells (HEPG2), as determined by metabolic activity (CellTiter-Blue assay). Data are presented as mean values ± SD; P = 0.0093 and P = 0.0027; unpaired two-tailed t test; n = 3 different experiments.

Article Snippet: CD14 was stained with Alexa Fluor 488 anti-human CD14 antibody (no. 367130; BioLegend) (1:100), CD9 was stained with Alexa Fluor 647 anti-human CD9 antibody (no. MCA469A647T; Bio-Rad) (1:1,000), TLR4 was stained with mouse anti-TLR4 antibody (no. NBP1-51697; R&D Systems) (3 μg/ml) and Alexa Fluor 647 goat anti-mouse IgG (H+L) secondary antibody (no. A-21235; Thermo Fisher) (1:500), and CD11b was stained with rabbit anti-CD11b antibody (no. 133357; Abcam) and Alexa Fluor 488 goat anti-rabbit IgG (H+L) secondary antibody.

Techniques: Incubation, Software, Labeling, Fluorescence, Two Tailed Test, Activity Assay, CtB Assay

Journal: Cell reports

Article Title: Single-Cell Analysis of the Muscle Stem Cell Hierarchy Identifies Heterotypic Communication Signals Involved in Skeletal Muscle Regeneration

doi: 10.1016/j.celrep.2020.02.067

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-Mouse CD9 (KMC8) , Fluidigm , Cat# 3158009B.

Techniques: Recombinant, Staining, Labeling, Software

Journal: Immunology

Article Title: The CD19/CD81 complex physically interacts with CD38 but is not required to induce proliferation in mouse B lymphocytes

doi: 10.1111/j.1365-2567.2012.03602.x

Figure Lengend Snippet: Co-capping of CD38 with CD63 and CD9. (a) Splenocytes from C57BL/6 mice were incubated with rabbit polyclonal anti-CD38 antibody for 30 min on ice, washed and reacted with a secondary anti-rabbit-Cy3 antibody for 1 hr at 37°. Counterstaining was performed at 4° with anti-CD63 (a) or CD9 (b) and DAPI was used for staining of nuclei. Confocal sections for the merged images of representative cells are shown. Coefficient of correlation (CC) for colocalization was calculated using imagej software. (b) The number of co-caps is presented as percentage of cells analyzed. Confocal sections for the merged images of representative cells are shown.

Article Snippet: Anti-CD81 (104907) and anti-CD19 (115513) were purchase from Biolegend (San Diego, CA), anti-B220-FITC (553088) and anti-CD9 (558749) were purchase from BD Pharmingen (San Diego, CA); and anti-CD63 (sc15363) from Santa Cruz Biotechnology, Inc (CA) and secondary antibodies anti-rabbit-Cy3 (Caltag Laboratories, Burlingame, CA; {"type":"entrez-nucleotide","attrs":{"text":"L42010","term_id":"804648","term_text":"L42010"}} L42010 ), anti-hamster-FITC (BD Pharmingen; 554011), anti-rabbit-horseradish peroxidase (HRP) (Thermo Scientific, Pittsburgh, PA; 1858415), anti-hamster-HRP (BD Pharmingen; 554012), anti-rat-HRP (ZyMax, San Francisco, CA; 81-9520) and anti-mouse-HRP (Thermo Scientific; 1858413).

Techniques: Incubation, Staining, Software

Journal: Journal of Neurovirology

Article Title: MicroRNA in neuroexosome as a potential biomarker for HIV-associated neurocognitive disorders

doi: 10.1007/s13365-024-01241-8

Figure Lengend Snippet: List of antibodies used in this study

Article Snippet: CD81 , Mouse , 17B1 , Immunoprecipitation , , − , FUJIFILM Wako.

Techniques: Western Blot, Immunoprecipitation

Journal: Journal of Neurovirology

Article Title: MicroRNA in neuroexosome as a potential biomarker for HIV-associated neurocognitive disorders

doi: 10.1007/s13365-024-01241-8

Figure Lengend Snippet: Characterization of total exosomes extracted from plasma. a Transmission electron micrographs of total exosomes derived from non-HIV controls. Scale bar, 100 nm. b Transmission electron micrographs of total exosomes derived from PLWH diagnosed with HAD. Scale bar, 100 nm. c NanoSight analysis of total exosomes derived from non-HIV controls. The black line indicates the means. Red indicates distribution. d NanoSight analysis of total exosomes derived from PLWH diagnosed with HAD. The black line indicates the means. Red indicates distribution. e Detection of the exosome markers CD9, CD81, and CD63 in total exosomes derived from non-HIV controls. Calnexin shows cell contamination

Article Snippet: CD81 , Mouse , 17B1 , Immunoprecipitation , , − , FUJIFILM Wako.

Techniques: Clinical Proteomics, Transmission Assay, Derivative Assay

Journal: Journal of Neurovirology

Article Title: MicroRNA in neuroexosome as a potential biomarker for HIV-associated neurocognitive disorders

doi: 10.1007/s13365-024-01241-8

Figure Lengend Snippet: CD81- and L1CAM-positive exosomes expressed the neuronal markers enolase-2 and miR-124–1. a CD9 and enolase-2 detection using western blotting. CD9 is a panexosome marker. Enolase-2 is a neuroexosome marker. T-E: total exosomes isolated from non-HIV controls, Control: exosomes immunoprecipitated by control IgG, CD81: exosomes immunoprecipitated by anti-CD81 antibody, L1CAM: exosomes immunoprecipitated by anti-L1CAM antibody, CD63: exosomes immunoprecipitated by anti-CD63 antibody. b Enolase-2 expression levels in total exosomes isolated from non-HIV controls ( n = 5), CD81-positive exosomes ( n = 5), L1CAM-positive exosomes ( n = 5), and CD63-positive exosomes ( n = 5). The intensity of the enolase-2 signal detected by western blotting was normalized to the intensity of the CD9 signal. * p < 0.05. c Expression of tissue-specific miRNAs in total exosomes ( n = 5), CD81-positive exosomes ( n = 5), L1CAM-positive exosomes ( n = 5), and CD63-positive exosomes ( n = 5). miR-122: liver marker, miR-124–1: neuronal marker, miR-125: intestinal organ marker. * p < 0.05

Article Snippet: CD81 , Mouse , 17B1 , Immunoprecipitation , , − , FUJIFILM Wako.

Techniques: Western Blot, Marker, Isolation, Control, Immunoprecipitation, Expressing