rabbit anti human antibody cd44 apc  (Thermo Fisher)

Journal: International Journal of Oncology

Article Title: Anisomycin inhibits angiogenesis in ovarian cancer by attenuating the molecular sponge effect of the lncRNA-Meg3/miR-421/PDGFRA axis

doi: 10.3892/ijo.2019.4887

Figure Lengend Snippet: Proportion of CD44 + /CD133 + cancer stem cells in ovarian cancer tissues of each patient by flow cytometry.

Article Snippet: The cell precipitates were collected and incubated with mouse anti-human CD133-FITC antibodies (1:100; cat. no. 11-1339-42; eBioscience; Thermo Fisher Scientific, Inc.) and rabbit anti-human CD44-APC antibodies (1:100; cat. no. 17-0441-82; eBioscience; Thermo Fisher Scientific, Inc.) at 4°C for 30 min. Next, CD44 + /CD133 + OCSCs were sorted from the sample by fluorescence-activated cell sorting.

Techniques: Flow Cytometry

Journal: International Journal of Oncology

Article Title: Anisomycin inhibits angiogenesis in ovarian cancer by attenuating the molecular sponge effect of the lncRNA-Meg3/miR-421/PDGFRA axis

doi: 10.3892/ijo.2019.4887

Figure Lengend Snippet: Anisomycin inhibits proliferation of CD44+/CD133+ HuOCSCs. (A) Proliferation of HuOCSCs, treated as indicated, was evaluated with the MTT assay. (B) Quantification and (C) representative plots from Annexin V/PI staining and flow cytometry analysis of HuOCSCs, treated as indicated. ** P<0.01 vs. non-treated cells (n=4). HuOCSCs, human ovarian cancer stem cells.

Article Snippet: The cell precipitates were collected and incubated with mouse anti-human CD133-FITC antibodies (1:100; cat. no. 11-1339-42; eBioscience; Thermo Fisher Scientific, Inc.) and rabbit anti-human CD44-APC antibodies (1:100; cat. no. 17-0441-82; eBioscience; Thermo Fisher Scientific, Inc.) at 4°C for 30 min. Next, CD44 + /CD133 + OCSCs were sorted from the sample by fluorescence-activated cell sorting.

Techniques: MTT Assay, Staining, Flow Cytometry

Journal: International Journal of Oncology

Article Title: Anisomycin inhibits angiogenesis in ovarian cancer by attenuating the molecular sponge effect of the lncRNA-Meg3/miR-421/PDGFRA axis

doi: 10.3892/ijo.2019.4887

Figure Lengend Snippet: Anisomycin inhibits HuOCSC invasion and angiogenesis in vitro . (A) A Transwell invasion assay was used to investigate the invasive properties of CD44 + /CD133 + HuOCSCs following anisomycin treatment (n=4). Magnification, ×100. (B) Representative images and (C) quantification of the ability of HUVECs to form 3-dimensional tubular structures in Matrigel following anisomycin treatment (n=4). Magnification, ×100. (D) Quantification and (E) representative images of the growth and development of intersegmental vessels and dorsal longitudinal anastomotic vessels in zebrafish embryos following anisomycin treatment (n=30). Magnification, ×40. ** P<0.01 vs. non-treated. HuOCSCs, human ovarian cancer stem cells; GFP, green fluorescent protein.

Article Snippet: The cell precipitates were collected and incubated with mouse anti-human CD133-FITC antibodies (1:100; cat. no. 11-1339-42; eBioscience; Thermo Fisher Scientific, Inc.) and rabbit anti-human CD44-APC antibodies (1:100; cat. no. 17-0441-82; eBioscience; Thermo Fisher Scientific, Inc.) at 4°C for 30 min. Next, CD44 + /CD133 + OCSCs were sorted from the sample by fluorescence-activated cell sorting.

Techniques: In Vitro, Transwell Invasion Assay

Journal: International Journal of Oncology

Article Title: Anisomycin inhibits angiogenesis in ovarian cancer by attenuating the molecular sponge effect of the lncRNA-Meg3/miR-421/PDGFRA axis

doi: 10.3892/ijo.2019.4887

Figure Lengend Snippet: Anisomycin inhibits the in vivo activity of CD44 + /CD133 + HuOCSCs. (A) Tumours formed by anisomycin-pre-treated and DMSO-pretreated HuOCSCs in nude mice. (B) Volume of tumours throughout the experiment. (C) Final tumour weights at 35 weeks. (D) H&E staining histopathology analysis of the xenografted tumours. Blood vessels are indicated by blue arrows. Magnification, ×200. (E) Immunofluorescence staining of the density of CD31/Flt-1 and FVIII/CD146 double-positive cells in the xenografted tumours. Magnification, ×200. ** P<0.01 vs. DMSO group (n=4). HuOCSCs, human ovarian cancer stem cells; Flt-1, fms related tyrosine kinase 1; FVIII, coagulation factor VIII.

Article Snippet: The cell precipitates were collected and incubated with mouse anti-human CD133-FITC antibodies (1:100; cat. no. 11-1339-42; eBioscience; Thermo Fisher Scientific, Inc.) and rabbit anti-human CD44-APC antibodies (1:100; cat. no. 17-0441-82; eBioscience; Thermo Fisher Scientific, Inc.) at 4°C for 30 min. Next, CD44 + /CD133 + OCSCs were sorted from the sample by fluorescence-activated cell sorting.

Techniques: In Vivo, Activity Assay, Staining, Histopathology, Immunofluorescence, Coagulation

Journal: International Journal of Oncology

Article Title: Anisomycin inhibits angiogenesis in ovarian cancer by attenuating the molecular sponge effect of the lncRNA-Meg3/miR-421/PDGFRA axis

doi: 10.3892/ijo.2019.4887

Figure Lengend Snippet: Anisomycin affects the expression of PDGFRs and Notch pathway components in ovarian cancer stem cells. (A) mRNA expression levels of key factors of the Notch pathway, PDGFRA and PDGFRB in ovarian cancer tissues with more and less CD44 + /CD133 + HuOCSCs. ** P<0.01. (B) mRNA expression levels of key factors of the Notch pathway, PDGFRA and PDGFRB in CD44 + /CD133 + HuOCSCs following anisomycin treatment. ** P<0.01 vs. non-treated (n=4). (C) Western blotting results from the experimental groups of panel A. (D) Western blotting results from the experimental groups of panel B. PDGFR, platelet-derived growth factor receptor; HuOCSCs, human ovarian cancer stem cells; DLL1, δ-like canonical Notch ligand 1; HES1, hes family bHLH transcription factor 1; RBPJ, recombination signal binding protein for immunoglobulin κ J region.

Article Snippet: The cell precipitates were collected and incubated with mouse anti-human CD133-FITC antibodies (1:100; cat. no. 11-1339-42; eBioscience; Thermo Fisher Scientific, Inc.) and rabbit anti-human CD44-APC antibodies (1:100; cat. no. 17-0441-82; eBioscience; Thermo Fisher Scientific, Inc.) at 4°C for 30 min. Next, CD44 + /CD133 + OCSCs were sorted from the sample by fluorescence-activated cell sorting.

Techniques: Expressing, Western Blot, Derivative Assay, Binding Assay