Journal: Stem Cells International

Article Title: Enhanced Expression of ABCB1 and Nrf2 in CD133-Positive Cancer Stem Cells Associates with Doxorubicin Resistance

doi: 10.1155/2020/8868849

Figure Lengend Snippet: The separation of HCT8 colorectal cancer cells into different subpopulations based on the expression of CD44 and CD133. (a) Representative dot plots of flow cytometry analysis show the purities of each subpopulation of isolated cells. Quantitative data in the dot plots are presented as the percentages of positive cells from three independent experiments. (b) Representative photos of morphological properties (upper) and MTT assay on cell growth (lower) at 24 hr after the initiation of culture. Data are presented as the mean ± SD from three independent experiments. (c, d) Representative histograms of flow cytometry analysis showed the expressions of CD44 (c) and CD133 (d) at baseline and 45 days after cell culture. The dotted vertical lines through histograms indicate the difference in the expression peaks between the baseline and at 45 days after culture. Quantitative data in the histograms are presented as the mean fluorescent intensity from three independent experiments.

Article Snippet: To verify the purity of each subpopulation, isolated cells were stained with PE-labelled mouse anti-human CD133 (clone: AC133) (Miltenyi Biotec) and FITC-labelled mouse anti-human CD44 (clone: DB105) (Miltenyi Biotec), according to the supplied protocols.

Techniques: Expressing, Flow Cytometry, Isolation, MTT Assay, Cell Culture

Journal: Stem Cells International

Article Title: Enhanced Expression of ABCB1 and Nrf2 in CD133-Positive Cancer Stem Cells Associates with Doxorubicin Resistance

doi: 10.1155/2020/8868849

Figure Lengend Snippet: DXR resistance of different subpopulations of cells. (a) MTT assay was done to evaluate the cytotoxicity of DXR. Data are expressed as the percentile of baseline (before DXR treatment) from three independent experiments. ∗ P < 0.01 vs. all other subpopulations. (b) Representative histograms of flow cytometry analysis show the accumulation of DXR in cells 24 hr after the treatment with 10 μ M DXR, in the absence or presence of 50 μ M verapamil and 200 μ M BSO. The dotted vertical lines through histograms indicated the mean levels of DXR accumulation in CD44 + CD133 + cells for comparing with other subpopulations of cells. The results were reproducible in three independent experiments. (c) Representative histograms of flow cytometry analysis show the expression of the ABCB1 or ABCG2 in different subpopulations of cells. Quantitative data in the histograms are presented as the mean fluorescent intensity from three independent experiments.

Article Snippet: To verify the purity of each subpopulation, isolated cells were stained with PE-labelled mouse anti-human CD133 (clone: AC133) (Miltenyi Biotec) and FITC-labelled mouse anti-human CD44 (clone: DB105) (Miltenyi Biotec), according to the supplied protocols.

Techniques: MTT Assay, Flow Cytometry, Expressing

Journal: Stem Cells International

Article Title: Enhanced Expression of ABCB1 and Nrf2 in CD133-Positive Cancer Stem Cells Associates with Doxorubicin Resistance

doi: 10.1155/2020/8868849

Figure Lengend Snippet: DXR resistance and nuclear DXR accumulation in CD44 + CD133 − and CD44 + CD133 + cells, in the absence or presence of drug efflux inhibitor. (a) MTT assay was done to compare the cytotoxicity of DXR in CD44 + CD133 − and CD44 + CD133 + cells, with or without the addition of 50 μ M verapamil. Data were expressed as a percent of baseline (before DXR treatment) from three independent experiments. ∗ P < 0.05 vs. CD44 + CD133 − cells. (b) Representative histograms of flow cytometry analysis showed the nuclear accumulation of DXR in cells 24 hr after the treatment by 10 μ M DXR, with or without the addition of 50 μ M verapamil. The results were reproducible in three independent experiments.

Article Snippet: To verify the purity of each subpopulation, isolated cells were stained with PE-labelled mouse anti-human CD133 (clone: AC133) (Miltenyi Biotec) and FITC-labelled mouse anti-human CD44 (clone: DB105) (Miltenyi Biotec), according to the supplied protocols.

Techniques: MTT Assay, Flow Cytometry

Journal: Stem Cells International

Article Title: Enhanced Expression of ABCB1 and Nrf2 in CD133-Positive Cancer Stem Cells Associates with Doxorubicin Resistance

doi: 10.1155/2020/8868849

Figure Lengend Snippet: Different expression of phosphorylated p38MAPK between CD44 + CD133 − and CD44 + CD133 + cells. Representative blots and semiquantitative data on the expression of phosphorylated p38MAPK and total p38MAPK in cells treated with 10 μ M DXR, in the absence or presence of 50 μ M verapamil. The quantitative data are normalized to total p38MAPK. Data are expressed as relative values to CD44 + CD133 − cells without DXR treatment and presented as the mean ± SD from three independent experiments.

Article Snippet: To verify the purity of each subpopulation, isolated cells were stained with PE-labelled mouse anti-human CD133 (clone: AC133) (Miltenyi Biotec) and FITC-labelled mouse anti-human CD44 (clone: DB105) (Miltenyi Biotec), according to the supplied protocols.

Techniques: Expressing

Journal: Stem Cells International

Article Title: Enhanced Expression of ABCB1 and Nrf2 in CD133-Positive Cancer Stem Cells Associates with Doxorubicin Resistance

doi: 10.1155/2020/8868849

Figure Lengend Snippet: Different antioxidant capacity between CD44 + CD133 − and CD44 + CD133 + cells. (a) Representative histograms of flow cytometry analysis show the intracellular ROS levels 24 hr after the treatment by 10 μ M DXR, in the absence or presence of 50 μ M verapamil. The results were reproducible in three independent experiments. (b) Representative blots and semiquantitative data on the expression of Nrf2 in cells treated with 10 μ M DXR, in the absence or presence of 50 μ M verapamil. The quantitative data are normalized to β -tubulin. Data are expressed as relative values to CD44 + CD133 − cells without DXR treatment and presented as the mean ± SD from three independent experiments.

Article Snippet: To verify the purity of each subpopulation, isolated cells were stained with PE-labelled mouse anti-human CD133 (clone: AC133) (Miltenyi Biotec) and FITC-labelled mouse anti-human CD44 (clone: DB105) (Miltenyi Biotec), according to the supplied protocols.

Techniques: Flow Cytometry, Expressing

Journal: Stem Cells International

Article Title: Enhanced Expression of ABCB1 and Nrf2 in CD133-Positive Cancer Stem Cells Associates with Doxorubicin Resistance

doi: 10.1155/2020/8868849

Figure Lengend Snippet: The effect of silencing CD133 expression on DXR resistance of CD44 + CD133 + cells. (a) Representative histograms of flow cytometry analysis on the expression of CD133 in CD44 + CD133 + cells after silencing by different dosages of targeted siRNA. Quantitative data in the histograms are presented as the mean fluorescent intensity from three independent experiments. (b) MTT assay was done to evaluate the cytotoxicity to DXR. Cells were treated with 5 nM siRNA for 48 hr followed by DXR treatment for another 48 hr. Data are expressed as a percent of baseline (before DXR treatment) from three independent experiments. ∗ P < 0.05 vs. CD44 + CD133 − cells. (c) Representative histograms of flow cytometry analysis on the expression of ABCB1 in cells after silencing by different dosages of targeted siRNA. Quantitative data in the histograms are presented as the mean fluorescent intensity from three independent experiments. (d) Representative histograms of flow cytometry analysis on the accumulation of DXR. Quantitative data in the histograms are presented as the mean fluorescent intensity from three independent experiments.

Article Snippet: To verify the purity of each subpopulation, isolated cells were stained with PE-labelled mouse anti-human CD133 (clone: AC133) (Miltenyi Biotec) and FITC-labelled mouse anti-human CD44 (clone: DB105) (Miltenyi Biotec), according to the supplied protocols.

Techniques: Expressing, Flow Cytometry, MTT Assay

Journal: Oncology Letters

Article Title: p53 positively regulates the expression of cancer stem cell marker CD133 in HCT116 colon cancer cells

doi: 10.3892/ol.2018.8619

Figure Lengend Snippet: Differential expression of the CSC surface marker CD133 in p53+/+ and p53−/− HCT116 cell lines. (A) Western blot analysis of p53 expression in HCT116 cell lines. (B) Flow cytometric analysis of CD133 and CD44 expression in p53+/+ and p53−/− HCT116 cell lines. (C) Quantification of flow cytometry results for CD133 and CD44 expression in the two HCT116 cell lines. *P<0.01, with comparisons indicated by brackets. (D) CD133 mRNA expression levels in p53+/+ and p53−/− HCT116 cell lines were detected by reverse transcription-quantitative polymerase chain reaction. *P<0.01 for p53+/+ vs. p53−/− HCT116 cells. (E) Western blot analysis of CD133 protein expression. CSC, cancer stem cell; PE, phycoerythrin; FITC, fluorescein isothiocyanate.

Article Snippet: Next, 10 µl of the CD133/1 (AC133)-fluorescein isothiocyanate (FITC) antibody (clone, 393C3, cat. no. 130-104-322) and 10 µl of the CD44-phycoerythrin (PE) antibody (clone DB105, cat. no. 130-098-108) (both from Miltenyi Biotec, GmbH, Bergisch, Germany) were added, mixed well and incubated for 10 min in the dark at 4°C.

Techniques: Expressing, Marker, Western Blot, Flow Cytometry, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Spontaneous development of Epstein-Barr Virus associated human lymphomas in a prostate cancer xenograft program

doi: 10.1371/journal.pone.0188228

Figure Lengend Snippet: A . Serially transplantable PDX tumour were depleted of mouse cells before labelling with human specific antibodies to CD44, CD45, CD19, CD56 and EPCaM. B . Table of percentage of cells expressing specific markers. LNCaP was used as a positive control for EPCaM expression and PC3 was used as a negative control for CD45, CD19 and CD56 expression. C . IHC of Chromogranin A expression in PDX lines.

Article Snippet: Cells harvested from xenografts were analysed for the expression of human CD44 (clone DB105; Miltenyi Biotec), human CD24 (clone 32D12; Miltenyi Biotec), human EpCAM (clone CD326; Miltenyi Biotech), human B lymphocyte antigen, CD19 (clone HIB19; eBioscience), human neural cell adhesion molecule, CD56 (clone AF12-7H3; Miltenyi Biotec) and human CD45 (clones H130 and 2D1; eBioscience) following mouse cell depletion (Miltenyi Biotech).

Techniques: Expressing, Positive Control, Negative Control

Journal: Disease Models & Mechanisms

Article Title: Extended passaging of the SKOV3 ovarian cancer cell line leads to two phenotypically different strains

doi: 10.1242/dmm.052451

Figure Lengend Snippet: Flow cytometry analysis of SKOV3 strains S1 and S2 was performed at different passages, from P2 to P50 after isolation by differential trypsinization. (A) Histograms display the expression of markers ESA, CD24, CD90 and CD73 at passages 5, 25 and 50. (B) Changes in marker expression are shown in histograms for passages 2, 25, 35 and 50. The markers analysed include ESA, CD44, CD24, CD90, CD73 and CD274.

Article Snippet: Cells were incubated with the following monoclonal antibodies: anti-ESA (CD326)-PE (Miltenyi Biotec, MACS, clone HEA-125, cat. no. 130-113-826, 1:50), CD44-APC (Miltenyi Biotec, MACS, clone DB105, cat. no. 130-113-893, 1:50), CD24-FITC [eBioscienceTM, InvitrogenTM, clone eBioSN3 (SN3 A5-2H10), cat. no. 15576356, 1:20], CD90-PE/Cy7 (BioLegend, clone 5E10, cat. no. 328123, 1:25), CD73-PE (BioLegend, clone AD2, cat. no. 344003, 1:25) and CD274-PE (BD Bioscience, clone MIH1, cat. no. 561787, 1:25).

Techniques: Flow Cytometry, Isolation, Expressing, Marker