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: Biology of reproduction

Article Title: Sperm-oocyte signaling: the role of IZUMO1R and CD9 in PTK2B activation and actin remodeling at the sperm binding site†.

doi: 10.1093/biolre/ioab048

Figure Lengend Snippet: Figure 1. Progression of PTK2B-PY402, IZUMO1R, and CD9 accumulation at sperm binding sites. Zona-free oocytes collected between 10 and 20 min post- insemination were prepared for immunofluorescence and labeled with anti-PTK2B-PY402 (green) and anti-IZUMO1R (red) (columns A-D) or with anti-PTK2B- PY402 and anti-CD9 (red) (columns E-H). Magnification is indicated by the bars, which represents 10 um.

Article Snippet: Expression of murine cd9 in oocytes null for mouse cd9 The mouse cd9 clone (# MG226288, ORIGENE (Rockville, MD)) was used to transfect E. coli (CopyCutter C400CH10, Lucigen Corporation (Middleton, WI)) and the sequence of the insert was confirmed.

Techniques: Binding Assay, Labeling

Journal: Biology of reproduction

Article Title: Sperm-oocyte signaling: the role of IZUMO1R and CD9 in PTK2B activation and actin remodeling at the sperm binding site†.

doi: 10.1093/biolre/ioab048

Figure Lengend Snippet: Figure 3. Effect of Izumo1r or Cd9 ablation on PTK2B recruitment at sperm binding sites. Oocytes collected from wt, Izumo1r−/−, or Cd9−/−females were incubated with capacitated sperm as above, fixed and labeled with anti-PTK2B (green). Magnification is indicated by the bars, which represents 10 um.

Article Snippet: Expression of murine cd9 in oocytes null for mouse cd9 The mouse cd9 clone (# MG226288, ORIGENE (Rockville, MD)) was used to transfect E. coli (CopyCutter C400CH10, Lucigen Corporation (Middleton, WI)) and the sequence of the insert was confirmed.

Techniques: Binding Assay, Incubation, Labeling

Journal: Biology of reproduction

Article Title: Sperm-oocyte signaling: the role of IZUMO1R and CD9 in PTK2B activation and actin remodeling at the sperm binding site†.

doi: 10.1093/biolre/ioab048

Figure Lengend Snippet: Figure 2. Changes in the distribution of PTK2B-PY402 and CD9 during early sperm incorporation. Zona-free wt oocytes were collected between 20 and 30 mpi and labelled with anti-PTK2B-PY402 (green) and anti-CD9 (red). Column A represents an “early” stage where the sperm has deformed the oocyte cortex. Column B demonstrates a sperm that is being drawn into the oocyte. Column C is a volume view produced by z-stack images of a different binding site that demonstrates the ring-like distribution of CD9 and the near absence of PTK2B-PY402 as seen from “above”. Magnification is indicated by the bar, which represents 10 um.

Article Snippet: Expression of murine cd9 in oocytes null for mouse cd9 The mouse cd9 clone (# MG226288, ORIGENE (Rockville, MD)) was used to transfect E. coli (CopyCutter C400CH10, Lucigen Corporation (Middleton, WI)) and the sequence of the insert was confirmed.

Techniques: Produced, Binding Assay

Journal: Biology of reproduction

Article Title: Sperm-oocyte signaling: the role of IZUMO1R and CD9 in PTK2B activation and actin remodeling at the sperm binding site†.

doi: 10.1093/biolre/ioab048

Figure Lengend Snippet: Figure 4. Effect of Izumo1r or Cd9 ablation on PTK2B phosphorylation at sperm binding sites. (A) Oocytes collected from wt, Izumo1r −/−, or Cd9 −/−females were incubated with capacitated sperm, fixed and labeled with anti-PTK2B-PY402 (green) and anti-CD9 (red). Magnification is indicated by the bars, which represents 10 um. (B) The mean relative fluorescence intensity of PTK2B-PY402 at sperm binding sites of wt (white) and Izumo1r−/−oocytes (gray) from Table 1. (C) Percentage of wt and Izumo1r−/−sperm binding sites with increasing levels of PTK2B-PY402 fluorescence intensity relative to adjacent cortex. (D) The mean relative fluorescence intensity of PTK2B-PY402 at sperm binding sites of wt (white) and Cd9−/−oocytes (gray) from Table 1. (E) Percentage of wt and Cd9−/−

Article Snippet: Expression of murine cd9 in oocytes null for mouse cd9 The mouse cd9 clone (# MG226288, ORIGENE (Rockville, MD)) was used to transfect E. coli (CopyCutter C400CH10, Lucigen Corporation (Middleton, WI)) and the sequence of the insert was confirmed.

Techniques: Phospho-proteomics, Binding Assay, Incubation, Labeling

Journal: Biology of reproduction

Article Title: Sperm-oocyte signaling: the role of IZUMO1R and CD9 in PTK2B activation and actin remodeling at the sperm binding site†.

doi: 10.1093/biolre/ioab048

Figure Lengend Snippet: Figure 6. Effect of Izumo1r or Cd9 ablation on f-actin accumulation at sperm binding sites. Zona-free oocytes from wt, Izumo1r−/−, or Cd9−/−females with bound sperm were fixed and labeled with alexa568-phalloidin (red) to detect f-actin. Columns A and B represent early stages of sperm-oocyte interaction. (C) Mean relative fluorescence intensity of f-actin at sperm binding sites of wt (white) and Izumo1r−/−oocytes (gray) from Table 2. (D) Histogram presentation of the percentage of wt and Izumo1r−/−sperm binding sites with increasing levels of PTK2B-PY402 fluorescence intensity relative to adjacent cortex. (E) The mean relative fluorescence intensity of f-actin at sperm binding sites of wt (white) and Cd9−/−oocytes (gray) from Table 2. (F) Percentage of wt and Cd9−/−sperm binding sites with increasing levels of PTK2B-PY402 fluorescence intensity relative to adjacent cortex. Magnification is indicated by the bar, which represents 10 um.

Article Snippet: Expression of murine cd9 in oocytes null for mouse cd9 The mouse cd9 clone (# MG226288, ORIGENE (Rockville, MD)) was used to transfect E. coli (CopyCutter C400CH10, Lucigen Corporation (Middleton, WI)) and the sequence of the insert was confirmed.

Techniques: Binding Assay, Labeling

Journal: Biology of reproduction

Article Title: Sperm-oocyte signaling: the role of IZUMO1R and CD9 in PTK2B activation and actin remodeling at the sperm binding site†.

doi: 10.1093/biolre/ioab048

Figure Lengend Snippet: Figure 5. Rescue of Cd9−/−oocytes by injected Cd9 mRNA, GV stage oocytes from Cd9−/−females were injected with mRNA encoding c-Fyn (c-Fyn inj), with buffer only (sham inj), or with Cd9 mRNA (Cd9 inj) as described in “Methods”. (A) Matured oocytes with bound sperm were fixed and labeled with anti-CD9 (red) and anti-PTK2B-PY402 (green). (B) Relative PTK2B-PY402 fluorescence intensity at sperm binding sites from c-Fyn-injected and Cd9-injected oocytes. (C) Mean PTK2B-PY402 fluorescence intensity at sperm binding sites from c-Fyn-injected and Cd9-injected oocytes ±SEM. (D) Histogram presentation of the percentage of sperm binding sites with increasing levels of PTK2B-PY402 fluorescence. Magnification is indicated by the bars, which represent 10 um. Data were obtained from 3 replicates, each from at least two females, totaling 16 oocytes in each group. The number of sperm binding sites examined included 89 from the c-Fyn-injected group and 151 from the cd9-injected group.

Article Snippet: Expression of murine cd9 in oocytes null for mouse cd9 The mouse cd9 clone (# MG226288, ORIGENE (Rockville, MD)) was used to transfect E. coli (CopyCutter C400CH10, Lucigen Corporation (Middleton, WI)) and the sequence of the insert was confirmed.

Techniques: Injection, Labeling, Binding Assay

Journal: Biology of reproduction

Article Title: Sperm-oocyte signaling: the role of IZUMO1R and CD9 in PTK2B activation and actin remodeling at the sperm binding site†.

doi: 10.1093/biolre/ioab048

Figure Lengend Snippet: Figure 1. Progression of PTK2B-PY402, IZUMO1R, and CD9 accumulation at sperm binding sites. Zona-free oocytes collected between 10 and 20 min post- insemination were prepared for immunofluorescence and labeled with anti-PTK2B-PY402 (green) and anti-IZUMO1R (red) (columns A-D) or with anti-PTK2B- PY402 and anti-CD9 (red) (columns E-H). Magnification is indicated by the bars, which represents 10 um.

Article Snippet: Expression of murine cd9 in oocytes null for mouse cd9 The mouse cd9 clone (# MG226288, ORIGENE (Rockville, MD)) was used to transfect E. coli (CopyCutter C400CH10, Lucigen Corporation (Middleton, WI)) and the sequence of the insert was confirmed.

Techniques: Binding Assay, Labeling

Journal: Biology of reproduction

Article Title: Sperm-oocyte signaling: the role of IZUMO1R and CD9 in PTK2B activation and actin remodeling at the sperm binding site†.

doi: 10.1093/biolre/ioab048

Figure Lengend Snippet: Figure 3. Effect of Izumo1r or Cd9 ablation on PTK2B recruitment at sperm binding sites. Oocytes collected from wt, Izumo1r−/−, or Cd9−/−females were incubated with capacitated sperm as above, fixed and labeled with anti-PTK2B (green). Magnification is indicated by the bars, which represents 10 um.

Article Snippet: Expression of murine cd9 in oocytes null for mouse cd9 The mouse cd9 clone (# MG226288, ORIGENE (Rockville, MD)) was used to transfect E. coli (CopyCutter C400CH10, Lucigen Corporation (Middleton, WI)) and the sequence of the insert was confirmed.

Techniques: Binding Assay, Incubation, Labeling

Journal: Biology of reproduction

Article Title: Sperm-oocyte signaling: the role of IZUMO1R and CD9 in PTK2B activation and actin remodeling at the sperm binding site†.

doi: 10.1093/biolre/ioab048

Figure Lengend Snippet: Figure 2. Changes in the distribution of PTK2B-PY402 and CD9 during early sperm incorporation. Zona-free wt oocytes were collected between 20 and 30 mpi and labelled with anti-PTK2B-PY402 (green) and anti-CD9 (red). Column A represents an “early” stage where the sperm has deformed the oocyte cortex. Column B demonstrates a sperm that is being drawn into the oocyte. Column C is a volume view produced by z-stack images of a different binding site that demonstrates the ring-like distribution of CD9 and the near absence of PTK2B-PY402 as seen from “above”. Magnification is indicated by the bar, which represents 10 um.

Article Snippet: Expression of murine cd9 in oocytes null for mouse cd9 The mouse cd9 clone (# MG226288, ORIGENE (Rockville, MD)) was used to transfect E. coli (CopyCutter C400CH10, Lucigen Corporation (Middleton, WI)) and the sequence of the insert was confirmed.

Techniques: Produced, Binding Assay

Journal: Biology of reproduction

Article Title: Sperm-oocyte signaling: the role of IZUMO1R and CD9 in PTK2B activation and actin remodeling at the sperm binding site†.

doi: 10.1093/biolre/ioab048

Figure Lengend Snippet: Figure 4. Effect of Izumo1r or Cd9 ablation on PTK2B phosphorylation at sperm binding sites. (A) Oocytes collected from wt, Izumo1r −/−, or Cd9 −/−females were incubated with capacitated sperm, fixed and labeled with anti-PTK2B-PY402 (green) and anti-CD9 (red). Magnification is indicated by the bars, which represents 10 um. (B) The mean relative fluorescence intensity of PTK2B-PY402 at sperm binding sites of wt (white) and Izumo1r−/−oocytes (gray) from Table 1. (C) Percentage of wt and Izumo1r−/−sperm binding sites with increasing levels of PTK2B-PY402 fluorescence intensity relative to adjacent cortex. (D) The mean relative fluorescence intensity of PTK2B-PY402 at sperm binding sites of wt (white) and Cd9−/−oocytes (gray) from Table 1. (E) Percentage of wt and Cd9−/−

Article Snippet: Expression of murine cd9 in oocytes null for mouse cd9 The mouse cd9 clone (# MG226288, ORIGENE (Rockville, MD)) was used to transfect E. coli (CopyCutter C400CH10, Lucigen Corporation (Middleton, WI)) and the sequence of the insert was confirmed.

Techniques: Phospho-proteomics, Binding Assay, Incubation, Labeling

Journal: Biology of reproduction

Article Title: Sperm-oocyte signaling: the role of IZUMO1R and CD9 in PTK2B activation and actin remodeling at the sperm binding site†.

doi: 10.1093/biolre/ioab048

Figure Lengend Snippet: Figure 6. Effect of Izumo1r or Cd9 ablation on f-actin accumulation at sperm binding sites. Zona-free oocytes from wt, Izumo1r−/−, or Cd9−/−females with bound sperm were fixed and labeled with alexa568-phalloidin (red) to detect f-actin. Columns A and B represent early stages of sperm-oocyte interaction. (C) Mean relative fluorescence intensity of f-actin at sperm binding sites of wt (white) and Izumo1r−/−oocytes (gray) from Table 2. (D) Histogram presentation of the percentage of wt and Izumo1r−/−sperm binding sites with increasing levels of PTK2B-PY402 fluorescence intensity relative to adjacent cortex. (E) The mean relative fluorescence intensity of f-actin at sperm binding sites of wt (white) and Cd9−/−oocytes (gray) from Table 2. (F) Percentage of wt and Cd9−/−sperm binding sites with increasing levels of PTK2B-PY402 fluorescence intensity relative to adjacent cortex. Magnification is indicated by the bar, which represents 10 um.

Article Snippet: Expression of murine cd9 in oocytes null for mouse cd9 The mouse cd9 clone (# MG226288, ORIGENE (Rockville, MD)) was used to transfect E. coli (CopyCutter C400CH10, Lucigen Corporation (Middleton, WI)) and the sequence of the insert was confirmed.

Techniques: Binding Assay, Labeling

Journal: Biology of reproduction

Article Title: Sperm-oocyte signaling: the role of IZUMO1R and CD9 in PTK2B activation and actin remodeling at the sperm binding site†.

doi: 10.1093/biolre/ioab048

Figure Lengend Snippet: Figure 5. Rescue of Cd9−/−oocytes by injected Cd9 mRNA, GV stage oocytes from Cd9−/−females were injected with mRNA encoding c-Fyn (c-Fyn inj), with buffer only (sham inj), or with Cd9 mRNA (Cd9 inj) as described in “Methods”. (A) Matured oocytes with bound sperm were fixed and labeled with anti-CD9 (red) and anti-PTK2B-PY402 (green). (B) Relative PTK2B-PY402 fluorescence intensity at sperm binding sites from c-Fyn-injected and Cd9-injected oocytes. (C) Mean PTK2B-PY402 fluorescence intensity at sperm binding sites from c-Fyn-injected and Cd9-injected oocytes ±SEM. (D) Histogram presentation of the percentage of sperm binding sites with increasing levels of PTK2B-PY402 fluorescence. Magnification is indicated by the bars, which represent 10 um. Data were obtained from 3 replicates, each from at least two females, totaling 16 oocytes in each group. The number of sperm binding sites examined included 89 from the c-Fyn-injected group and 151 from the cd9-injected group.

Article Snippet: Expression of murine cd9 in oocytes null for mouse cd9 The mouse cd9 clone (# MG226288, ORIGENE (Rockville, MD)) was used to transfect E. coli (CopyCutter C400CH10, Lucigen Corporation (Middleton, WI)) and the sequence of the insert was confirmed.

Techniques: Injection, Labeling, Binding 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: Biomaterials

Article Title: Cleaved SPP1-rich extracellular vesicles from osteoclasts promote bone regeneration via TGFβ1/SMAD3 signaling.

doi: 10.1016/j.biomaterials.2023.122367

Figure Lengend Snippet: Fig. 5. Osteoclast EVs contained osteogenic proteins. A) The classification of proteins in OC-EVs according to the cellular components. B) The STRING analysis of the top 20 % of proteins (extracellular components) from OC-EVs for protein-protein interactions. C) mRNA expression of SPP1,CST3, ITGβ1, and OMD during the osteoclastic differentiation. D) Confocal microscopic images of OCs stained with CD9 (green), SPP1 (Red), CST3 (Red), and DAPI (blue) at day 6. The scale bar represents 100 μm. Data were expressed as means ± standard deviation (SD). *p < 0.05; **p < 0.001; ***p < 0.0001; ****p < 0.0001. One-way ANOVA with Dunnett’s post hoc test (n = 6). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Then cells were incubated with primary antibodies targeting osteopontin (22952-1-AP, Protein Tech), cystatin C (12245-1-AP, Protein Tech), and CD9 (MAB5218 R&D System) followed by washing and further labeling with secondary antibodies conjugated with fluorescent dyes.

Techniques: Protein-Protein interactions, Expressing, Staining, Standard Deviation