Journal: PLoS ONE

Article Title: N-Glycoproteome of E14.Tg2a Mouse Embryonic Stem Cells

doi: 10.1371/journal.pone.0055722

Figure Lengend Snippet: Differentially identified mESCs CD proteins in our mES E14.Tg2a N-glycoproteome and mES D3 surface proteome [43] .

Article Snippet: The anti-CD230 antibody was from Abcam, and antibodies against Cd133 and Cd90 were from BD Biosciences.

Techniques:

Journal: Sleep & Breathing = Schlaf & Atmung

Article Title: Altered profile of circulating endothelial progenitor cells in obstructive sleep apnea

doi: 10.1007/s11325-012-0781-4

Figure Lengend Snippet: Association between sleep parameters, endothelial progenitor cells, and advanced glycation end-products

Article Snippet: Then, 100 μL of citrated blood was incubated with monoclonal antibodies against human kinase insert domain receptor (KDR) (1:10, R&D Biosystems; FITC-conjugated), CD133 (1:10, Miltenyi; PE-conjugated), CD34 (1:10, Becton Dickinson; APC-conjugated), and CD45 (1:10, Beckman Coulter; PC7-conjugated) for 30 min. After incubation, blood was lysed and fixed by OptiLyse® C solution (Beckman Coulter).

Techniques:

Journal: Sleep & Breathing = Schlaf & Atmung

Article Title: Altered profile of circulating endothelial progenitor cells in obstructive sleep apnea

doi: 10.1007/s11325-012-0781-4

Figure Lengend Snippet: Characteristics of recruited subjects

Article Snippet: Then, 100 μL of citrated blood was incubated with monoclonal antibodies against human kinase insert domain receptor (KDR) (1:10, R&D Biosystems; FITC-conjugated), CD133 (1:10, Miltenyi; PE-conjugated), CD34 (1:10, Becton Dickinson; APC-conjugated), and CD45 (1:10, Beckman Coulter; PC7-conjugated) for 30 min. After incubation, blood was lysed and fixed by OptiLyse® C solution (Beckman Coulter).

Techniques:

Journal: Clinical and Translational Medicine

Article Title: APE1 shRNA‐loaded cancer stem cell‐derived extracellular vesicles reverse Erlotinib resistance in non‐small cell lung cancer via the IL‐6/STAT3 signalling

doi: 10.1002/ctm2.876

Figure Lengend Snippet: Isolation and characterization of CSC‐EVs as well as their internalization by NSCLC cells. (A) Expression of CSC surface markers (ABCG2 and CD133) in HCC827P‐CSCs and HCC827R‐CSCs, detected by flow cytometry. (B) Cell viability of HCC827P‐CSCs and HCC827R‐CSCs treated with different concentrations of Erlotinib, detected by CCK‐8 assay. (C) Size distribution of EVs derived from HCC827P‐CSCs and HCC827R‐CSCs measured by NTA. (D) Images of TEM observation of HCC827P‐CSCs and HCC827R‐CSCs. (E) Protein bands of EV surface markers (CD63, TSG101 and Calnexin) in Western blot of EVs (labelled E) and cell lysate (labelled CL) in HCC827P‐CSC‐EVs and HCC827R‐CSC‐EVs. (F) Images photographed in fluorescence microscopic observation of green fluorescence PKH67 in HCC827P and PC9P cells following co‐culture with PKH67‐labelled HCC827P‐CSC‐EVs and HCC827R‐CSC‐EVs (DAPI‐labelled nuclei are in blue). * p < .05, compared to HCC827P‐CSCs. Each cell experiment was conducted in triplicate. CSC, cancer stem cell; EV, extracellular vesicle; NSCLC, non‐small cell lung cancer; TEM, transmission electron microscope

Article Snippet: HCC827P, PC9P, HCC827R and PC9R cells were subjected to flow cytometric analysis to obtain CD133 + ABCG2 + CSCs (labelled HCC827P‐CSCs, PC9P‐CSCs, HCC827R‐CSCs and PC9R‐CSCs, respectively), with corresponding antibodies against CSC markers (anti‐CD133, ab216323, Abcam, Cambridge, UK; anti‐ABCG2, ab229193 Abcam) used in the analysis.

Techniques: Isolation, Expressing, Flow Cytometry, CCK-8 Assay, Derivative Assay, Western Blot, Fluorescence, Co-Culture Assay, Transmission Assay, Microscopy

Journal: Clinical and Translational Medicine

Article Title: APE1 shRNA‐loaded cancer stem cell‐derived extracellular vesicles reverse Erlotinib resistance in non‐small cell lung cancer via the IL‐6/STAT3 signalling

doi: 10.1002/ctm2.876

Figure Lengend Snippet: Effect of HCC827P‐CSC‐EVs and HCC827R‐CSC‐EVs on the resistance of NSCLC cells to Erlotinib. (A) Expression of Erlotinib resistance‐related genes MDR1, MRP, LRP and ABCG2 in HCC827P and PC9P cells following co‐culture with HCC827P‐CSC‐EVs or HCC827R‐CSC‐EVs, detected by qRT‐PCR and Western blot. (B) Viability of HCC827P and PC9P cells co‐cultured with HCC827P‐CSC‐EVs or HCC827R‐CSC‐EVs and further treated with different concentrations of Erlotinib, detected by CCK‐8 assay. Quantification of the migration (C) and invasion (D) of Erlotinib (5 μM)‐treated HCC827P and PC9P cells in response to co‐culture with HCC827P‐CSC‐EVs or HCC827R‐CSC‐EVs, observed by Transwell assay. (E) Apoptosis of Erlotinib (5 μM)‐treated HCC827P and PC9P cells in response to co‐culture with HCC827P‐CSC‐EVs or HCC827R‐CSC‐EVs, detected by flow cytometry. (F) Protein expression of anti‐apoptotic Bcl‐2 and pro‐apoptotic Bax and cleaved caspase‐3 in Western blot of Erlotinib (5 μM)‐treated HCC827P and PC9P cells in response to co‐culture with HCC827P‐CSC‐EVs or HCC827R‐CSC‐EVs. * p < .05. Each cell experiment was conducted in triplicate. CSC, cancer stem cell; EV, extracellular vesicle; NSCLC, non‐small cell lung cancer

Article Snippet: HCC827P, PC9P, HCC827R and PC9R cells were subjected to flow cytometric analysis to obtain CD133 + ABCG2 + CSCs (labelled HCC827P‐CSCs, PC9P‐CSCs, HCC827R‐CSCs and PC9R‐CSCs, respectively), with corresponding antibodies against CSC markers (anti‐CD133, ab216323, Abcam, Cambridge, UK; anti‐ABCG2, ab229193 Abcam) used in the analysis.

Techniques: Expressing, Co-Culture Assay, Quantitative RT-PCR, Western Blot, Cell Culture, CCK-8 Assay, Migration, Transwell Assay, Flow Cytometry

Journal: Clinical and Translational Medicine

Article Title: APE1 shRNA‐loaded cancer stem cell‐derived extracellular vesicles reverse Erlotinib resistance in non‐small cell lung cancer via the IL‐6/STAT3 signalling

doi: 10.1002/ctm2.876

Figure Lengend Snippet: APE1 affects the Erlotinib resistance of NSCLC cells through mediating the IL‐6/STAT3 signalling. (A) The protein expression of APE1 and p‐STAT3/STAT3 in HCC827R and PC9R cells treated by Erlotinib alone or in combination with shAPE1, measured by the Western blot. (B) The content of IL‐6 in the supernatant of HCC827R and PC9R cells treated by Erlotinib alone or in combination with shAPE1, detected by ELISA. (C) Cell viability in HCC827R and PC9R cells treated by Erlotinib alone or in combination with shAPE1, detected by CCK‐8 assay. Quantification of the migration (D) and invasion (E) in HCC827R and PC9R cells treated by Erlotinib alone or in combination with shAPE1, observed in the Transwell assay. (F) The apoptosis of HCC827R and PC9R cells treated by Erlotinib alone or in combination with shAPE1, tested by flow cytometry. (G) The protein expression of Bcl‐2, Bax and cleaved caspase‐3 in HCC827R and PC9R cells treated by Erlotinib alone or in combination with shAPE1, measured by the Western blot. (H) The mRNA and protein expression of MDR1, MRP, LRP and ABCG2 in CC827R and PC9R cells treated by Erlotinib alone or in combination with shAPE1, measured by qRT‐PCR and Western blot. * p < .05. Each cell experiment was conducted in triplicate. APE1, apurinic endonuclease 1; IL‐6, interleukin‐6; NSCLC, non‐small cell lung cancer

Article Snippet: HCC827P, PC9P, HCC827R and PC9R cells were subjected to flow cytometric analysis to obtain CD133 + ABCG2 + CSCs (labelled HCC827P‐CSCs, PC9P‐CSCs, HCC827R‐CSCs and PC9R‐CSCs, respectively), with corresponding antibodies against CSC markers (anti‐CD133, ab216323, Abcam, Cambridge, UK; anti‐ABCG2, ab229193 Abcam) used in the analysis.

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Migration, Transwell Assay, Flow Cytometry, Quantitative RT-PCR

Journal: Clinical and Translational Medicine

Article Title: APE1 shRNA‐loaded cancer stem cell‐derived extracellular vesicles reverse Erlotinib resistance in non‐small cell lung cancer via the IL‐6/STAT3 signalling

doi: 10.1002/ctm2.876

Figure Lengend Snippet: Effects of HCC827R‐CSC‐EVs loaded with APE1 shRNA on the Erlotinib resistance of NSCLC cells in vitro. (A) The enrichment of APEX1 (APE1) in EVs of different origins, analysed using the ExoRBase database. (B) The mRNA expression of APE1 in HCC827P‐CSCs, HCC827R‐CSCs, HCC827P‐CSC‐EVs and HCC827R‐CSC‐EVs determined by qRT‐PCR assay. (C) The mRNA expression of APE1 in HCC827R‐CSCs and HCC827R‐CSC‐EVs in response to shAPE1 treatment, determined by qRT‐PCR assay. (D) The mRNA expression of APE1 in HCC827P and PC9P cells in response to co‐culture with HCC827R‐CSCs EVs shAPE1 determined by qRT‐PCR assay. (E) Western blot measurement of the protein expression of p‐STAT3/STAT3 in HCC827P and PC9P cells in response to co‐culture with HCC827R‐CSCs EVs shAPE1. (F) ELISA detection of the content of IL‐6 in the supernatant of HCC827P and PC9P cells in response to co‐culture with HCC827R‐CSCs EVs shAPE1. (G) Cell viability in HCC827P and PC9P cells in response to co‐culture with HCC827R‐CSCs EVs shAPE1 and further treatment with Erlotinib, detected by CCK‐8 assay. Quantification of the migration (H) and invasion (I) in HCC827P and PC9P cells in response to co‐culture with HCC827R‐CSCs EVs shAPE1 and further treatment with Erlotinib (5 μM), detected by the Transwell assay. (J) Cell apoptosis in HCC827P and PC9P cells in response to co‐culture with HCC827R‐CSCs EVs shAPE1 and further treatment with Erlotinib (5 μM), detected by flow cytometry. (K) Protein expression of anti‐apoptotic Bcl‐2 and pro‐apoptotic Bax and cleaved caspase‐3 in the Western blot of Erlotinib (5 μM)‐treated HCC827P and PC9P cells in response to co‐culture with HCC827R‐CSCs EVs shAPE1. (L) The expression of MDR1, MRP, LRP and ABCG2 in Erlotinib (5 μM)‐treated HCC827P and PC9P cells in response to co‐culture with HCC827R‐CSCs EVs shAPE1 measured by qRT‐PCR and the Western blot. * p < .05. Each cell experiment was conducted in triplicate. APE1, apurinic endonuclease 1; CSC, cancer stem cell; EV, extracellular vesicle; IL‐6, interleukin‐6; NSCLC, non‐small cell lung cancer; shRNA, short hairpin RNA

Article Snippet: HCC827P, PC9P, HCC827R and PC9R cells were subjected to flow cytometric analysis to obtain CD133 + ABCG2 + CSCs (labelled HCC827P‐CSCs, PC9P‐CSCs, HCC827R‐CSCs and PC9R‐CSCs, respectively), with corresponding antibodies against CSC markers (anti‐CD133, ab216323, Abcam, Cambridge, UK; anti‐ABCG2, ab229193 Abcam) used in the analysis.

Techniques: shRNA, In Vitro, Expressing, Quantitative RT-PCR, Co-Culture Assay, Western Blot, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Migration, Transwell Assay, Flow Cytometry

Journal: Clinical and Translational Medicine

Article Title: APE1 shRNA‐loaded cancer stem cell‐derived extracellular vesicles reverse Erlotinib resistance in non‐small cell lung cancer via the IL‐6/STAT3 signalling

doi: 10.1002/ctm2.876

Figure Lengend Snippet: Effects of shAPE1‐loaded HCC827R‐CSC‐EVs on the Erlotinib resistance of NSCLC cells in nude mice. Nude mice were treated with Erlotinib, Erlotinib + HCC827R‐CSC‐EVs or Erlotinib + HCC827R‐CSC‐EVs shAPE1. (A) The volume of tumour xenografts at different time points. (B) The weight of isolated tumour xenografts in mice. (C) ELISA detection of IL‐6 protein levels in tumour tissues of mice. (D) The Western blot measurement of the protein expression of APE1, p‐STAT3 and STAT3 in tumour tissues of mice. (E) The Western blot measurement of anti‐apoptotic Bcl‐2 and pro‐apoptotic Bax and cleaved caspase‐3 in tumour tissue of mice. (F) The expression of MDR1, MRP, LRP and ABCG2 in tumour tissues of mice measured by qRT‐PCR and Western blot. * p < .05. n = 8. APE1, apurinic endonuclease 1; CSC, cancer stem cell; EV, extracellular vesicle; IL‐6, interleukin‐6; NSCLC, non‐small cell lung cancer

Article Snippet: HCC827P, PC9P, HCC827R and PC9R cells were subjected to flow cytometric analysis to obtain CD133 + ABCG2 + CSCs (labelled HCC827P‐CSCs, PC9P‐CSCs, HCC827R‐CSCs and PC9R‐CSCs, respectively), with corresponding antibodies against CSC markers (anti‐CD133, ab216323, Abcam, Cambridge, UK; anti‐ABCG2, ab229193 Abcam) used in the analysis.

Techniques: Isolation, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Quantitative RT-PCR

Journal: American Journal of Translational Research

Article Title: Enhanced penetration of exogenous EPCs into brains of APP/PS1 transgenic mice

doi:

Figure Lengend Snippet: Identification of EPC biomarkers by immunofluorescence. A. Mouse MNCs from bone marrow double labeling by FITC-CD34+ cells and Dylight594-CD133+ cells (magnified 200 X). B. The cells labeled by FITC and Dylight594, separately (magnified 200 X).

Article Snippet: Adherent cells were digested into a single-cell suspension, blocked for 30 min and then incubated with antibodies against either mouse CD133 (Millipore, Cat #: MAB4310) or mouse CD34 (Abcam, Cat #: 2150-1) for 30 min at 4°C.

Techniques: Immunofluorescence, Labeling

Journal: American Journal of Translational Research

Article Title: Enhanced penetration of exogenous EPCs into brains of APP/PS1 transgenic mice

doi:

Figure Lengend Snippet: Flow cytometric analysis of surface markers of EPCs differentiated from mouse bone marrow MNCs. Data are presented as the mean ± SD, n=3 (*p< 0.05). A. The expression of CD133 and CD34 were separately detected using FL1-H and FL2-H, respectively. Control represents negative control with primary antibody incubation. B. The percentage of positive cells, corresponding data in A.

Article Snippet: Adherent cells were digested into a single-cell suspension, blocked for 30 min and then incubated with antibodies against either mouse CD133 (Millipore, Cat #: MAB4310) or mouse CD34 (Abcam, Cat #: 2150-1) for 30 min at 4°C.

Techniques: Expressing, Control, Negative Control, Incubation

Journal: BMC Developmental Biology

Article Title: Vascularization of primary and secondary ossification centres in the human growth plate

doi: 10.1186/s12861-014-0036-7

Figure Lengend Snippet: CD34, α-SMA, CD133, CD90, CD105, CXCR4, VEGFR1, VEGFR2 expression in growth plate and in the surrounding GOR (n = 23). An entire digit section was scanned by TissueFAXS (overview) including uncalcified cartilage (C), area of ongoing calcification (*), joint cavity (JC), surrounding soft tissue (ST), articular cartilage (AC), Groove of Ranvier (GOR), POC, and SOC. Immature endothelial cells in POCs expressed CD34 (A) , abluminal coverage with α-SMA + smooth muscle cells was found on vessels in newly formed bone but not in POCs and SOCs (B) . Single CD133 precursor cells were found in POCs in close proximity to newly formed vessels (C) , mesenchymal progenitor cells expressing CD90 (D) and CD105 (E) co-localized with CD34 + endothelial cells in POCs. CXCR4+ cells (F) could be localized in POCs. VEGFR1 (G) is expressed on endothelial cells of vessels in newly formed bone, while VEGFR2 (H) is expressed on immature CD34 + endothelial cells in POCs. The groove of Ranvier (GOR) (J) showed intense vascularisation (K) with mesenchymal progenitors expressing CD105 (L) . A representative isotype control was included (I) . Large arrowheads indicate the area of the insert ( scale bar equals 50 μm), small arrowheads indicate specific cells.

Article Snippet: Sections of paraffin embedded tissues were treated with horse serum (CD133, CD34, CD45, CXCR4, RANK and OPG) or goat serum albumin (RANKL, DLX5, RUNX2) for 30 min at room temperature and subsequently incubated with the primary antibodies against CD133 (2.5 μg/ml mouse monoclonal antibody (mAb), Miltenyi Biotec, Bergisch Gladbach, Germany; 1.3 μg/ml goat polyclonal antibody (pAb), Santa Cruz Biotechnology, Santa Cruz, CA), CD34 (2.5 μg/ml mouse mAb, Becton Dickinson, San Jose, CA, or 0.5 μg/ml mouse mAb, Immunotech, Marseilles, France), CXCR4 (2.5 μg/ml mouse mAb, PharMingen, San Diego, CA), VEGFR-2 (2.5 μg/ml mouse mAb, Santa Cruz Biotechnology, Cell Signalling Technology, Danvers, MA), CD45 (3.5 μg/ml mouse mAb, Becton Dickinson), CD90 (2 μg/ml mouse mAb, Novus Biologicals, LLC, Littleton, CO), CD105 (2 μg/ml mouse mAb, Labvision/Neomarkers, Freemont, CA), RANKL (2 μg/ml rabbit mAb, Santa Cruz Biotechnology), RANK (2 μg/ml goat pAb, Santa Cruz Biotechnology), RUNX2 (2 μg/ml rabbit mAb, Oncogene, Cambridge, MA), DLX5 (2 μg/ml rabbit mAb, Chemicon, Billerica, MA), and OPG (2 μg/ml mouse mAb, R&D, Minneapolis, MN) at 4°C overnight.

Techniques: Expressing, Control

Journal: BMC Developmental Biology

Article Title: Vascularization of primary and secondary ossification centres in the human growth plate

doi: 10.1186/s12861-014-0036-7

Figure Lengend Snippet: Morphometric analysis: Number of CD34, CD45, CD133, CXCR4, RANK, RANKL, OPG, RUNX-2 and DLX-5 positive cells/mm 2 in the resting zone (RZ), proliferating zone (PZ), hypertrophic zone (HZ) and calcification zone (CZ) of the growth plate were counted in 10 fields of 1 mm 2 (n = 12). Statistically significant differences are indicated as * p < 0.05, ** p < 0.005, *** p < 0.001.

Article Snippet: Sections of paraffin embedded tissues were treated with horse serum (CD133, CD34, CD45, CXCR4, RANK and OPG) or goat serum albumin (RANKL, DLX5, RUNX2) for 30 min at room temperature and subsequently incubated with the primary antibodies against CD133 (2.5 μg/ml mouse monoclonal antibody (mAb), Miltenyi Biotec, Bergisch Gladbach, Germany; 1.3 μg/ml goat polyclonal antibody (pAb), Santa Cruz Biotechnology, Santa Cruz, CA), CD34 (2.5 μg/ml mouse mAb, Becton Dickinson, San Jose, CA, or 0.5 μg/ml mouse mAb, Immunotech, Marseilles, France), CXCR4 (2.5 μg/ml mouse mAb, PharMingen, San Diego, CA), VEGFR-2 (2.5 μg/ml mouse mAb, Santa Cruz Biotechnology, Cell Signalling Technology, Danvers, MA), CD45 (3.5 μg/ml mouse mAb, Becton Dickinson), CD90 (2 μg/ml mouse mAb, Novus Biologicals, LLC, Littleton, CO), CD105 (2 μg/ml mouse mAb, Labvision/Neomarkers, Freemont, CA), RANKL (2 μg/ml rabbit mAb, Santa Cruz Biotechnology), RANK (2 μg/ml goat pAb, Santa Cruz Biotechnology), RUNX2 (2 μg/ml rabbit mAb, Oncogene, Cambridge, MA), DLX5 (2 μg/ml rabbit mAb, Chemicon, Billerica, MA), and OPG (2 μg/ml mouse mAb, R&D, Minneapolis, MN) at 4°C overnight.

Techniques:

Journal: BMC Developmental Biology

Article Title: Vascularization of primary and secondary ossification centres in the human growth plate

doi: 10.1186/s12861-014-0036-7

Figure Lengend Snippet: mRNA levels of CD 34, CD31 and CD133 were quantified (n = 8) using RT-qPCR and expressed as numbers of molecules by 1,000 molecules of GAPDH (NMby1000GAPDH). Adherent cells isolated from dissected growth plates were cultured and analyzed separately. Results are given as mean values with standard deviation.

Article Snippet: Sections of paraffin embedded tissues were treated with horse serum (CD133, CD34, CD45, CXCR4, RANK and OPG) or goat serum albumin (RANKL, DLX5, RUNX2) for 30 min at room temperature and subsequently incubated with the primary antibodies against CD133 (2.5 μg/ml mouse monoclonal antibody (mAb), Miltenyi Biotec, Bergisch Gladbach, Germany; 1.3 μg/ml goat polyclonal antibody (pAb), Santa Cruz Biotechnology, Santa Cruz, CA), CD34 (2.5 μg/ml mouse mAb, Becton Dickinson, San Jose, CA, or 0.5 μg/ml mouse mAb, Immunotech, Marseilles, France), CXCR4 (2.5 μg/ml mouse mAb, PharMingen, San Diego, CA), VEGFR-2 (2.5 μg/ml mouse mAb, Santa Cruz Biotechnology, Cell Signalling Technology, Danvers, MA), CD45 (3.5 μg/ml mouse mAb, Becton Dickinson), CD90 (2 μg/ml mouse mAb, Novus Biologicals, LLC, Littleton, CO), CD105 (2 μg/ml mouse mAb, Labvision/Neomarkers, Freemont, CA), RANKL (2 μg/ml rabbit mAb, Santa Cruz Biotechnology), RANK (2 μg/ml goat pAb, Santa Cruz Biotechnology), RUNX2 (2 μg/ml rabbit mAb, Oncogene, Cambridge, MA), DLX5 (2 μg/ml rabbit mAb, Chemicon, Billerica, MA), and OPG (2 μg/ml mouse mAb, R&D, Minneapolis, MN) at 4°C overnight.

Techniques: Quantitative RT-PCR, Isolation, Cell Culture, Standard Deviation

Journal: PLoS ONE

Article Title: Sensitivity of Human Intrahepatic Cholangiocarcinoma Subtypes to Chemotherapeutics and Molecular Targeted Agents: A Study on Primary Cell Cultures

doi: 10.1371/journal.pone.0142124

Figure Lengend Snippet: Sequences of primer pairs (sense and antisense, respectively) used for amplifying the genes of interest (GOI) and the internal reference gene (GAPDH) used for their nor Primer designed by the PROBEFINDER software ( https://www.roche-applied-science.com/sis/rtpcr/upl/index.jsp ).

Article Snippet: Cells were rinsed twice with PBS buffer for 2 min, blocked and incubated 1 hour with the following antibodies: Vimentin mouse monoclonal IgG1 (sc-32322, Santa Cruz Biotechnology), α-SMA mouse monoclonal IgG1(M0851, Dako), E-Cadherin mouse monoclonal IgG1 (sc-21791, Santa Cruz Biotechnology), CD326/EpCAM mouse monoclonal IgG1 (sc-59782, Santa Cruz Biotechnology), CD133/PROM1 mouse monoclonal IgG1 (TA309943, Origene), LGR5 rabbit polyclonal IgG (TA301323,Origene), cytokeratin-19 (CK-19) sc-6278 CK-19 mouse monoclonal IgG2a (Santa Cruz Biotechnology), Interleukin 6 (IL6) mouse monoclonal IgG2a (ab9324, Abcam) at room temperature.

Techniques: Software, Amplification

Journal: PLoS ONE

Article Title: Sensitivity of Human Intrahepatic Cholangiocarcinoma Subtypes to Chemotherapeutics and Molecular Targeted Agents: A Study on Primary Cell Cultures

doi: 10.1371/journal.pone.0142124

Figure Lengend Snippet: (A) Immunohistochemical and immunofluorescence analyses of mixed- and mucin-IHCCA primary cultures for Vimentin, α-SMA, E-Cadherin and the “epithelial” cancer stem cell markers CD133+, EpCAM+, LGR5+ and for IL6. A diffuse positivity for mesenchymal markers (Vimentin, α-SMA) and IL6 was observed while E-Cadherin was virtually negative and less than 5% cells where positive for the “epithelial” cancer stem cell markers. Representative experiment of N = 12 independent staining performed in separate primary cultures. (B) Flow cytometry analyses of primary cultures of mixed- and mucin-IHCCA (20–30 passages), labeled with anti-CD13, anti-CD90, anti-EpCAM, anti-CD133, and anti-LGR5 antibodies. Bar graphs and representative plots. Cells positive for CD13 and CD90 largely predominated with respect to CD133, EpCAM and LGR5. CD13+ cells predominated in mixed-IHCCA with respect to mucin-IHCCA, while the opposite was found for CD90+ cells. Mean ± SD of N = 18 independent experiments. * = p< 0.05 vs mucin-IHCCA; & = p< 0.01 vs mixed-IHCCA.

Article Snippet: Cells were rinsed twice with PBS buffer for 2 min, blocked and incubated 1 hour with the following antibodies: Vimentin mouse monoclonal IgG1 (sc-32322, Santa Cruz Biotechnology), α-SMA mouse monoclonal IgG1(M0851, Dako), E-Cadherin mouse monoclonal IgG1 (sc-21791, Santa Cruz Biotechnology), CD326/EpCAM mouse monoclonal IgG1 (sc-59782, Santa Cruz Biotechnology), CD133/PROM1 mouse monoclonal IgG1 (TA309943, Origene), LGR5 rabbit polyclonal IgG (TA301323,Origene), cytokeratin-19 (CK-19) sc-6278 CK-19 mouse monoclonal IgG2a (Santa Cruz Biotechnology), Interleukin 6 (IL6) mouse monoclonal IgG2a (ab9324, Abcam) at room temperature.

Techniques: Immunohistochemical staining, Immunofluorescence, Staining, Flow Cytometry, Labeling

Journal: PLoS ONE

Article Title: Sensitivity of Human Intrahepatic Cholangiocarcinoma Subtypes to Chemotherapeutics and Molecular Targeted Agents: A Study on Primary Cell Cultures

doi: 10.1371/journal.pone.0142124

Figure Lengend Snippet: RT-PCR analysis of Vimentin and cancer stem cell surface markers.

Article Snippet: Cells were rinsed twice with PBS buffer for 2 min, blocked and incubated 1 hour with the following antibodies: Vimentin mouse monoclonal IgG1 (sc-32322, Santa Cruz Biotechnology), α-SMA mouse monoclonal IgG1(M0851, Dako), E-Cadherin mouse monoclonal IgG1 (sc-21791, Santa Cruz Biotechnology), CD326/EpCAM mouse monoclonal IgG1 (sc-59782, Santa Cruz Biotechnology), CD133/PROM1 mouse monoclonal IgG1 (TA309943, Origene), LGR5 rabbit polyclonal IgG (TA301323,Origene), cytokeratin-19 (CK-19) sc-6278 CK-19 mouse monoclonal IgG2a (Santa Cruz Biotechnology), Interleukin 6 (IL6) mouse monoclonal IgG2a (ab9324, Abcam) at room temperature.

Techniques: