CD133 Antibody Search Results


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  • 86
    Miltenyi Biotec cd133
    <t>CD19−CD133+</t> MCL cells demonstrate increased self-renewal, compared to CD19+CD133− MCL cells. (A) Diagram of the self-renewal assay. (B) CAFC analysis comparing bulk, CD133− and CD133+ MCL cells. (*p
    Cd133, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd133/product/Miltenyi Biotec
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd133 - by Bioz Stars, 2021-05
    86/100 stars
      Buy from Supplier

    cd133  (Abcam)
    86
    Abcam cd133
    Reduced miR-181b expression promotes CSC properties in NSCLC. A549/DDP and A549 cells were transfected with miR-181b mimics, miR-181b inhibitors, or the control. a , b The number of tumourspheres were counted, and the morphology was observed under a light microscope. c <t>CD133</t> + cells were analysed in A549/DDP and A549 spheres by flow cytometry. d The mRNA levels of KLF4, SOX2, NANOG, CD133, and ALDH were measured by qPCR. Bars represent 200 μm for the low-power lens and 50 μm for the high-power lens. Data are presented as the mean ± SD. * p
    Cd133, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd133/product/Abcam
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd133 - by Bioz Stars, 2021-05
    86/100 stars
      Buy from Supplier

    86
    Miltenyi Biotec anti cd133
    Molecular effects of cisplatin on <t>CD133‐Kd</t> cells. (A): Signaling pathways over‐represented by the 102 genes differentially expressed in shPROM1 in respect to CD133 + control cells after cisplatin treatment. (A) Panther analysis showed the enrichment of different pathways. The most represented pathways were PDGF signaling, Alzheimer, DNA replication, Wnt, and E‐cadherin related pathways (underlined in the figure). (B): Funrich confirmation of the biological pathways associated with genes disregulated in CD133‐Kd cisplatin treated cells (shPROM1 and shPROM2 cells). Columns represent the percentage of genes involved in each biological pathway.
    Anti Cd133, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd133/product/Miltenyi Biotec
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cd133 - by Bioz Stars, 2021-05
    86/100 stars
      Buy from Supplier

    86
    Abcam anti cd133
    The inhibitor of hVps34 can block SGK3 activity and suppress liver CSC expansion after prolonged treatment of HCC cells with PI3K inhibitors. a Huh7 and MHCC97H cells were treated with VPS34-IN1 for 24 h, and cell lysates were subjected to western blot analysis with the indicated antibodies. b Spheroid formation assay of MHCC97H cells treated with VPS34-IN1 (top), the statistical results of the tumour spheroids ( > 50 μm) were calculated based on 3 independent experiments (bottom). Scale bars, 200 μm. c Immunofluorescence images of Nanog (red) in MHCC7H cells after treatment with VPS34-IN1 or dimethyl sulphoxide DMSO for 24 h. Nuclei were stained with DAPI (blue). Scale bars, 10 μm. d The proportion of <t>CD133+</t> cells in Huh7 cells was evaluated by flow cytometric assay. e Treatment of MHCC97H cells with VPS34-IN1 for 24 h inhibited the overexpression of CD133 and Nanog induced by treatment with ZSTK474, detected by RT-PCR. * P
    Anti Cd133, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd133/product/Abcam
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cd133 - by Bioz Stars, 2021-05
    86/100 stars
      Buy from Supplier

    N/A
    Produced in rabbits immunized with a synthetic peptide corresponding to the C terminus of the Mouse PROM1 and purified by antigen affinity chromatography
      Buy from Supplier


    N/A
    PROM1 Antibody is a Rabbit Polyclonal antibody against PROM1 This gene encodes a pentaspan transmembrane glycoprotein The protein localizes to membrane protrusions and is often expressed on adult stem cells
      Buy from Supplier

    Image Search Results


    CD19−CD133+ MCL cells demonstrate increased self-renewal, compared to CD19+CD133− MCL cells. (A) Diagram of the self-renewal assay. (B) CAFC analysis comparing bulk, CD133− and CD133+ MCL cells. (*p

    Journal: PLoS ONE

    Article Title: Cobblestone-Area Forming Cells Derived from Patients with Mantle Cell Lymphoma Are Enriched for CD133+ Tumor-Initiating Cells

    doi: 10.1371/journal.pone.0091042

    Figure Lengend Snippet: CD19−CD133+ MCL cells demonstrate increased self-renewal, compared to CD19+CD133− MCL cells. (A) Diagram of the self-renewal assay. (B) CAFC analysis comparing bulk, CD133− and CD133+ MCL cells. (*p

    Article Snippet: Tonsil tissue (control) and tumor tissue were stained with H & E, antibodies specific for human CD3, CD5, CD23, CD19, CD20, CD45, CD79a, CD133, Cyclin D1, Pax5 and FISH analysis for the t(11;14) translocation. (TIFF) Click here for additional data file.

    Techniques:

    CD19−CD133+ MCL cells demonstrate increased quiescence. (A) A representative dot plot of Ki-67 stained MCL cell subpopulations (UPN1). (B) Bar graph summarizing the data derived from 6 patients (*p

    Journal: PLoS ONE

    Article Title: Cobblestone-Area Forming Cells Derived from Patients with Mantle Cell Lymphoma Are Enriched for CD133+ Tumor-Initiating Cells

    doi: 10.1371/journal.pone.0091042

    Figure Lengend Snippet: CD19−CD133+ MCL cells demonstrate increased quiescence. (A) A representative dot plot of Ki-67 stained MCL cell subpopulations (UPN1). (B) Bar graph summarizing the data derived from 6 patients (*p

    Article Snippet: Tonsil tissue (control) and tumor tissue were stained with H & E, antibodies specific for human CD3, CD5, CD23, CD19, CD20, CD45, CD79a, CD133, Cyclin D1, Pax5 and FISH analysis for the t(11;14) translocation. (TIFF) Click here for additional data file.

    Techniques: Staining, Derivative Assay

    Expression of of self-renewal and B-cell differentiation associated transcription factors. qRT-PCR was conducted on RNA isolated from UCB CD5+ B-cells, CD19+CD133− and CD19−CD133+ subsets of MCL cells.

    Journal: PLoS ONE

    Article Title: Cobblestone-Area Forming Cells Derived from Patients with Mantle Cell Lymphoma Are Enriched for CD133+ Tumor-Initiating Cells

    doi: 10.1371/journal.pone.0091042

    Figure Lengend Snippet: Expression of of self-renewal and B-cell differentiation associated transcription factors. qRT-PCR was conducted on RNA isolated from UCB CD5+ B-cells, CD19+CD133− and CD19−CD133+ subsets of MCL cells.

    Article Snippet: Tonsil tissue (control) and tumor tissue were stained with H & E, antibodies specific for human CD3, CD5, CD23, CD19, CD20, CD45, CD79a, CD133, Cyclin D1, Pax5 and FISH analysis for the t(11;14) translocation. (TIFF) Click here for additional data file.

    Techniques: Expressing, Cell Differentiation, Quantitative RT-PCR, Isolation

    CD19−CD133+ MCL cells demonstrate increased drug resistance. (D) Sensitivity of different subpopulations of MCL to Bortezomib (15 nM), doxorubicin (50 nM) and fludarabine (15 µg/ml) was evaluated in suspension assay and CAFC assay. (A) MCL subpopulations were cultured alone in the presence or absence of drug for 48 hrs then evaluated for cell viability by 7-AAD staining and FACS analysis *(p

    Journal: PLoS ONE

    Article Title: Cobblestone-Area Forming Cells Derived from Patients with Mantle Cell Lymphoma Are Enriched for CD133+ Tumor-Initiating Cells

    doi: 10.1371/journal.pone.0091042

    Figure Lengend Snippet: CD19−CD133+ MCL cells demonstrate increased drug resistance. (D) Sensitivity of different subpopulations of MCL to Bortezomib (15 nM), doxorubicin (50 nM) and fludarabine (15 µg/ml) was evaluated in suspension assay and CAFC assay. (A) MCL subpopulations were cultured alone in the presence or absence of drug for 48 hrs then evaluated for cell viability by 7-AAD staining and FACS analysis *(p

    Article Snippet: Tonsil tissue (control) and tumor tissue were stained with H & E, antibodies specific for human CD3, CD5, CD23, CD19, CD20, CD45, CD79a, CD133, Cyclin D1, Pax5 and FISH analysis for the t(11;14) translocation. (TIFF) Click here for additional data file.

    Techniques: Cell Culture, Staining, FACS

    Only CD19−CD133+ MCL engraft NOD/SCID mice and recapitulate the disease. (A) Diagram of serial transplantation scheme. (B) Representative survival curve of NOD/SCID mice injected with the various MCL subpopulations (UPN3). (C) Representative hCD45 engraftment profile of bone marrow (BM) and spleen (SP) from mice injected with either CD19+CD133− or CD19−CD133+ subpopulation (5,000 cells/mouse).

    Journal: PLoS ONE

    Article Title: Cobblestone-Area Forming Cells Derived from Patients with Mantle Cell Lymphoma Are Enriched for CD133+ Tumor-Initiating Cells

    doi: 10.1371/journal.pone.0091042

    Figure Lengend Snippet: Only CD19−CD133+ MCL engraft NOD/SCID mice and recapitulate the disease. (A) Diagram of serial transplantation scheme. (B) Representative survival curve of NOD/SCID mice injected with the various MCL subpopulations (UPN3). (C) Representative hCD45 engraftment profile of bone marrow (BM) and spleen (SP) from mice injected with either CD19+CD133− or CD19−CD133+ subpopulation (5,000 cells/mouse).

    Article Snippet: Tonsil tissue (control) and tumor tissue were stained with H & E, antibodies specific for human CD3, CD5, CD23, CD19, CD20, CD45, CD79a, CD133, Cyclin D1, Pax5 and FISH analysis for the t(11;14) translocation. (TIFF) Click here for additional data file.

    Techniques: Mouse Assay, Transplantation Assay, Injection

    Reduced miR-181b expression promotes CSC properties in NSCLC. A549/DDP and A549 cells were transfected with miR-181b mimics, miR-181b inhibitors, or the control. a , b The number of tumourspheres were counted, and the morphology was observed under a light microscope. c CD133 + cells were analysed in A549/DDP and A549 spheres by flow cytometry. d The mRNA levels of KLF4, SOX2, NANOG, CD133, and ALDH were measured by qPCR. Bars represent 200 μm for the low-power lens and 50 μm for the high-power lens. Data are presented as the mean ± SD. * p

    Journal: Stem Cell Research & Therapy

    Article Title: miR-181b/Notch2 overcome chemoresistance by regulating cancer stem cell-like properties in NSCLC

    doi: 10.1186/s13287-018-1072-1

    Figure Lengend Snippet: Reduced miR-181b expression promotes CSC properties in NSCLC. A549/DDP and A549 cells were transfected with miR-181b mimics, miR-181b inhibitors, or the control. a , b The number of tumourspheres were counted, and the morphology was observed under a light microscope. c CD133 + cells were analysed in A549/DDP and A549 spheres by flow cytometry. d The mRNA levels of KLF4, SOX2, NANOG, CD133, and ALDH were measured by qPCR. Bars represent 200 μm for the low-power lens and 50 μm for the high-power lens. Data are presented as the mean ± SD. * p

    Article Snippet: The slides were incubated with Notch2 (1:200 dilution, Abclonal), CD133 (1:200 dilution, JKSJ-orb372326), and SOX2 (1:100 dilution, Abcam-ab92494) primary antibodies, stained using DAB, and counterstained using haematoxylin.

    Techniques: Expressing, Transfection, Light Microscopy, Flow Cytometry, Cytometry, Real-time Polymerase Chain Reaction

    Overexpression of miR-181b inhibits CSC characteristics in vivo. Mice were treated with different doses of A549/DDP sphere cells transfected with miR-181b agomir or miR-181b-NC agomir. Representative images illustrating tumour growth ( a ) and tumour volume ( b ). c The mRNA level of CD133 was determined by qPCR. d Tumour growth and volume were evaluated after treatment with DDP (3.0 mg kg −1 body weight; i.p., thrice) or PBS (pH 7.4; i.p., thrice). e Immunostaining images showing Notch2 expression. The mRNA levels of Notch2 and miR-181b were determined by qPCR. Mice were treated with A549/DDP sphere cells and then with DAPT or the control. f Images show tumour growth and tumour volume. g , h The mRNA levels of CD133 and Notch2 were measured by qPCR. i PARP and cleaved PARP levels were also ascertained by Western blotting after the indicated treatment. Data are presented as the mean ± SD. * p

    Journal: Stem Cell Research & Therapy

    Article Title: miR-181b/Notch2 overcome chemoresistance by regulating cancer stem cell-like properties in NSCLC

    doi: 10.1186/s13287-018-1072-1

    Figure Lengend Snippet: Overexpression of miR-181b inhibits CSC characteristics in vivo. Mice were treated with different doses of A549/DDP sphere cells transfected with miR-181b agomir or miR-181b-NC agomir. Representative images illustrating tumour growth ( a ) and tumour volume ( b ). c The mRNA level of CD133 was determined by qPCR. d Tumour growth and volume were evaluated after treatment with DDP (3.0 mg kg −1 body weight; i.p., thrice) or PBS (pH 7.4; i.p., thrice). e Immunostaining images showing Notch2 expression. The mRNA levels of Notch2 and miR-181b were determined by qPCR. Mice were treated with A549/DDP sphere cells and then with DAPT or the control. f Images show tumour growth and tumour volume. g , h The mRNA levels of CD133 and Notch2 were measured by qPCR. i PARP and cleaved PARP levels were also ascertained by Western blotting after the indicated treatment. Data are presented as the mean ± SD. * p

    Article Snippet: The slides were incubated with Notch2 (1:200 dilution, Abclonal), CD133 (1:200 dilution, JKSJ-orb372326), and SOX2 (1:100 dilution, Abcam-ab92494) primary antibodies, stained using DAB, and counterstained using haematoxylin.

    Techniques: Over Expression, In Vivo, Mouse Assay, Transfection, Real-time Polymerase Chain Reaction, Immunostaining, Expressing, Western Blot

    miR-181b suppresses Notch2 signalling to inhibit CSC traits. a Venn diagrams show the number of genes identified as potential targets of miR-181b in three predictive programs: TargetScan, miRanda, and miRBD. Notch2 expression in adherent A549/DDP and A549 cells or cell spheres was tested by Western blotting. b Relative luciferase activity was evaluated after wild-type or mutant 3′-UTR reporter plasmids were co-transfected with pGLE-miR-181b or miR-181b-NC in H1299 cells. c A549 cells were transfected with miR-181b inhibitors or the control and then treated with si-Notch2 or negative control. Notch2 expression was analysed by Western blotting. d The number of tumourspheres was counted, and the morphology was observed under a light microscope. e The IC50 value was determined by CCK assay with different concentrations of cisplatin. The apoptotic percentage ( f ) and CD133 + cells ( g ) were evaluated by flow cytometry. h The mRNA levels of KLF4, SOX2, NANOG, CD133, and ALDH were measured by qPCR. i Western blotting showed Notch2, NICD2, HES1, and HEY1 expression after A549/DDP and A549 cells were transfected with miR-181b mimics, miR-181b inhibitors, or the control. Bars represent 200 μm for low-power lens and 50 μm for high-power lens. Data are presented as the mean ± SD. * p

    Journal: Stem Cell Research & Therapy

    Article Title: miR-181b/Notch2 overcome chemoresistance by regulating cancer stem cell-like properties in NSCLC

    doi: 10.1186/s13287-018-1072-1

    Figure Lengend Snippet: miR-181b suppresses Notch2 signalling to inhibit CSC traits. a Venn diagrams show the number of genes identified as potential targets of miR-181b in three predictive programs: TargetScan, miRanda, and miRBD. Notch2 expression in adherent A549/DDP and A549 cells or cell spheres was tested by Western blotting. b Relative luciferase activity was evaluated after wild-type or mutant 3′-UTR reporter plasmids were co-transfected with pGLE-miR-181b or miR-181b-NC in H1299 cells. c A549 cells were transfected with miR-181b inhibitors or the control and then treated with si-Notch2 or negative control. Notch2 expression was analysed by Western blotting. d The number of tumourspheres was counted, and the morphology was observed under a light microscope. e The IC50 value was determined by CCK assay with different concentrations of cisplatin. The apoptotic percentage ( f ) and CD133 + cells ( g ) were evaluated by flow cytometry. h The mRNA levels of KLF4, SOX2, NANOG, CD133, and ALDH were measured by qPCR. i Western blotting showed Notch2, NICD2, HES1, and HEY1 expression after A549/DDP and A549 cells were transfected with miR-181b mimics, miR-181b inhibitors, or the control. Bars represent 200 μm for low-power lens and 50 μm for high-power lens. Data are presented as the mean ± SD. * p

    Article Snippet: The slides were incubated with Notch2 (1:200 dilution, Abclonal), CD133 (1:200 dilution, JKSJ-orb372326), and SOX2 (1:100 dilution, Abcam-ab92494) primary antibodies, stained using DAB, and counterstained using haematoxylin.

    Techniques: Expressing, Western Blot, Luciferase, Activity Assay, Mutagenesis, Transfection, Negative Control, Light Microscopy, Flow Cytometry, Cytometry, Real-time Polymerase Chain Reaction

    Molecular effects of cisplatin on CD133‐Kd cells. (A): Signaling pathways over‐represented by the 102 genes differentially expressed in shPROM1 in respect to CD133 + control cells after cisplatin treatment. (A) Panther analysis showed the enrichment of different pathways. The most represented pathways were PDGF signaling, Alzheimer, DNA replication, Wnt, and E‐cadherin related pathways (underlined in the figure). (B): Funrich confirmation of the biological pathways associated with genes disregulated in CD133‐Kd cisplatin treated cells (shPROM1 and shPROM2 cells). Columns represent the percentage of genes involved in each biological pathway.

    Journal: Stem Cells Translational Medicine

    Article Title: Role of CD133 Molecule in Wnt Response and Renal Repair

    doi: 10.1002/sctm.17-0158

    Figure Lengend Snippet: Molecular effects of cisplatin on CD133‐Kd cells. (A): Signaling pathways over‐represented by the 102 genes differentially expressed in shPROM1 in respect to CD133 + control cells after cisplatin treatment. (A) Panther analysis showed the enrichment of different pathways. The most represented pathways were PDGF signaling, Alzheimer, DNA replication, Wnt, and E‐cadherin related pathways (underlined in the figure). (B): Funrich confirmation of the biological pathways associated with genes disregulated in CD133‐Kd cisplatin treated cells (shPROM1 and shPROM2 cells). Columns represent the percentage of genes involved in each biological pathway.

    Article Snippet: For Western blot analysis, anti‐CD133 (AC133 epitope) (Miltenyi Biotech), anti‐actin (Santa Cruz Biotechnology, Dallas, TX, USA), anti‐β‐catenin (R & D Systems, Minneapolis, MN, USA), anti‐E‐cadherin (BD Biosciences, Franklin Lakes, NJ, USA) and anti GSK3 alpha and beta (Abcam, Cambridge, UK) Abs were used.

    Techniques:

    Impaired activation of Wnt signaling in CD133‐Kd lines. (A, B): Quantitative real‐time PCR showing the expression of WNT4 upon cisplatin induced damage (A) or stimulation with CHIR99021 (B) by CD133 + (GFP) and CD133‐Kd (shPROM1 and shPROM2) cell lines. Data represent the mean ± SD of five different experiments and are normalized to GAPDH and to GFP. One way analysis of variance (ANOVA) was performed: *, p

    Journal: Stem Cells Translational Medicine

    Article Title: Role of CD133 Molecule in Wnt Response and Renal Repair

    doi: 10.1002/sctm.17-0158

    Figure Lengend Snippet: Impaired activation of Wnt signaling in CD133‐Kd lines. (A, B): Quantitative real‐time PCR showing the expression of WNT4 upon cisplatin induced damage (A) or stimulation with CHIR99021 (B) by CD133 + (GFP) and CD133‐Kd (shPROM1 and shPROM2) cell lines. Data represent the mean ± SD of five different experiments and are normalized to GAPDH and to GFP. One way analysis of variance (ANOVA) was performed: *, p

    Article Snippet: For Western blot analysis, anti‐CD133 (AC133 epitope) (Miltenyi Biotech), anti‐actin (Santa Cruz Biotechnology, Dallas, TX, USA), anti‐β‐catenin (R & D Systems, Minneapolis, MN, USA), anti‐E‐cadherin (BD Biosciences, Franklin Lakes, NJ, USA) and anti GSK3 alpha and beta (Abcam, Cambridge, UK) Abs were used.

    Techniques: Activation Assay, Real-time Polymerase Chain Reaction, Expressing

    Impaired proliferation and spheroid generation of CD133‐Kd cells after damage. (A): Proliferation of CD133 + (GFP) and CD133‐Kd (shPROM1 and shPROM2) cells was evaluated in the early phase of damage (CIS 48H) and during recovery phase (RECOVERY) by the incorporation of BrdU. Data are expressed as mean ± SD of three different experiments normalized to CTL and to one. One way analysis of variance (ANOVA) was performed: $ , p

    Journal: Stem Cells Translational Medicine

    Article Title: Role of CD133 Molecule in Wnt Response and Renal Repair

    doi: 10.1002/sctm.17-0158

    Figure Lengend Snippet: Impaired proliferation and spheroid generation of CD133‐Kd cells after damage. (A): Proliferation of CD133 + (GFP) and CD133‐Kd (shPROM1 and shPROM2) cells was evaluated in the early phase of damage (CIS 48H) and during recovery phase (RECOVERY) by the incorporation of BrdU. Data are expressed as mean ± SD of three different experiments normalized to CTL and to one. One way analysis of variance (ANOVA) was performed: $ , p

    Article Snippet: For Western blot analysis, anti‐CD133 (AC133 epitope) (Miltenyi Biotech), anti‐actin (Santa Cruz Biotechnology, Dallas, TX, USA), anti‐β‐catenin (R & D Systems, Minneapolis, MN, USA), anti‐E‐cadherin (BD Biosciences, Franklin Lakes, NJ, USA) and anti GSK3 alpha and beta (Abcam, Cambridge, UK) Abs were used.

    Techniques: CTL Assay

    Involvement of CD133 in cell senescence. (A): Representative micrograph of CD133 + (GFP) and CD133‐Kd (shPROM1 and shPROM2) cells stained for β‐galactosidase (β‐gal). Original magnification: ×200. The number of β‐gal positive cells in CD133 + and CD133‐Kd lines was evaluated as the percentage of β‐gal positive and negative cells/field in at least 10 fields per experiment (B) . Data are expressed as mean ± SD of three different experiments. (C): T/S ratio analysis expressed as mean ± SD of T/S ratio normalized to GFP and to one, of eight different experiments. One way analysis of variance was performed: *, p

    Journal: Stem Cells Translational Medicine

    Article Title: Role of CD133 Molecule in Wnt Response and Renal Repair

    doi: 10.1002/sctm.17-0158

    Figure Lengend Snippet: Involvement of CD133 in cell senescence. (A): Representative micrograph of CD133 + (GFP) and CD133‐Kd (shPROM1 and shPROM2) cells stained for β‐galactosidase (β‐gal). Original magnification: ×200. The number of β‐gal positive cells in CD133 + and CD133‐Kd lines was evaluated as the percentage of β‐gal positive and negative cells/field in at least 10 fields per experiment (B) . Data are expressed as mean ± SD of three different experiments. (C): T/S ratio analysis expressed as mean ± SD of T/S ratio normalized to GFP and to one, of eight different experiments. One way analysis of variance was performed: *, p

    Article Snippet: For Western blot analysis, anti‐CD133 (AC133 epitope) (Miltenyi Biotech), anti‐actin (Santa Cruz Biotechnology, Dallas, TX, USA), anti‐β‐catenin (R & D Systems, Minneapolis, MN, USA), anti‐E‐cadherin (BD Biosciences, Franklin Lakes, NJ, USA) and anti GSK3 alpha and beta (Abcam, Cambridge, UK) Abs were used.

    Techniques: Staining

    CD133‐Kd generation. The silencing of CD133 antigen in different cell lines was assessed by Western blot, quantitative real‐time PCR (qRT‐PCR) and cytofluorimetric analysis. (A): Representative Western blots of control GFP and different CD133‐Kd lines (shPROM1 and shPROM2). (B): Quantification of CD133 expression as evaluated by Western blot analysis. (C): Quantification of CD133 mRNA expression, normalized to GAPDH and to GFP, as evaluated by qRT‐PCR analysis. (D): Representative cytofluorimetric analysis and quantification as geometric mean of GFP and CD133‐Kd cell lines. The dark gray filled area represents the binding of CD133 mAb, while the overlaying black line represents the isotypic control. (B–D) Represent the mean ± SD of 12 different cell lines generated in the study and are normalized to GFP and to one. One way analysis of variance was performed: **, p

    Journal: Stem Cells Translational Medicine

    Article Title: Role of CD133 Molecule in Wnt Response and Renal Repair

    doi: 10.1002/sctm.17-0158

    Figure Lengend Snippet: CD133‐Kd generation. The silencing of CD133 antigen in different cell lines was assessed by Western blot, quantitative real‐time PCR (qRT‐PCR) and cytofluorimetric analysis. (A): Representative Western blots of control GFP and different CD133‐Kd lines (shPROM1 and shPROM2). (B): Quantification of CD133 expression as evaluated by Western blot analysis. (C): Quantification of CD133 mRNA expression, normalized to GAPDH and to GFP, as evaluated by qRT‐PCR analysis. (D): Representative cytofluorimetric analysis and quantification as geometric mean of GFP and CD133‐Kd cell lines. The dark gray filled area represents the binding of CD133 mAb, while the overlaying black line represents the isotypic control. (B–D) Represent the mean ± SD of 12 different cell lines generated in the study and are normalized to GFP and to one. One way analysis of variance was performed: **, p

    Article Snippet: For Western blot analysis, anti‐CD133 (AC133 epitope) (Miltenyi Biotech), anti‐actin (Santa Cruz Biotechnology, Dallas, TX, USA), anti‐β‐catenin (R & D Systems, Minneapolis, MN, USA), anti‐E‐cadherin (BD Biosciences, Franklin Lakes, NJ, USA) and anti GSK3 alpha and beta (Abcam, Cambridge, UK) Abs were used.

    Techniques: Western Blot, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Binding Assay, Generated

    Effect of cisplatin on the phenotype of CD133 + cells. (A): Regulated progenitor genes (green) and mesenchymal genes (red) in CD133 + cells subjected to cisplatin damage, by RNA sequencing. Data show the mean log ratio of three different lines and are presented as descending expression intensity. (B, C): Validation of gene modulation (PAX2, VCAM, FOXD9, SOX9, and CD133), normalized to GAPDH and to CTL, as evaluated by quantitative real‐time PCR analysis. Data are expressed as mean ± SD of at least three different experiments normalized to CTL and to one, t test or One way analysis of variance (ANOVA) (for CD133) was performed: *, p

    Journal: Stem Cells Translational Medicine

    Article Title: Role of CD133 Molecule in Wnt Response and Renal Repair

    doi: 10.1002/sctm.17-0158

    Figure Lengend Snippet: Effect of cisplatin on the phenotype of CD133 + cells. (A): Regulated progenitor genes (green) and mesenchymal genes (red) in CD133 + cells subjected to cisplatin damage, by RNA sequencing. Data show the mean log ratio of three different lines and are presented as descending expression intensity. (B, C): Validation of gene modulation (PAX2, VCAM, FOXD9, SOX9, and CD133), normalized to GAPDH and to CTL, as evaluated by quantitative real‐time PCR analysis. Data are expressed as mean ± SD of at least three different experiments normalized to CTL and to one, t test or One way analysis of variance (ANOVA) (for CD133) was performed: *, p

    Article Snippet: For Western blot analysis, anti‐CD133 (AC133 epitope) (Miltenyi Biotech), anti‐actin (Santa Cruz Biotechnology, Dallas, TX, USA), anti‐β‐catenin (R & D Systems, Minneapolis, MN, USA), anti‐E‐cadherin (BD Biosciences, Franklin Lakes, NJ, USA) and anti GSK3 alpha and beta (Abcam, Cambridge, UK) Abs were used.

    Techniques: RNA Sequencing Assay, Expressing, CTL Assay, Real-time Polymerase Chain Reaction

    The inhibitor of hVps34 can block SGK3 activity and suppress liver CSC expansion after prolonged treatment of HCC cells with PI3K inhibitors. a Huh7 and MHCC97H cells were treated with VPS34-IN1 for 24 h, and cell lysates were subjected to western blot analysis with the indicated antibodies. b Spheroid formation assay of MHCC97H cells treated with VPS34-IN1 (top), the statistical results of the tumour spheroids ( > 50 μm) were calculated based on 3 independent experiments (bottom). Scale bars, 200 μm. c Immunofluorescence images of Nanog (red) in MHCC7H cells after treatment with VPS34-IN1 or dimethyl sulphoxide DMSO for 24 h. Nuclei were stained with DAPI (blue). Scale bars, 10 μm. d The proportion of CD133+ cells in Huh7 cells was evaluated by flow cytometric assay. e Treatment of MHCC97H cells with VPS34-IN1 for 24 h inhibited the overexpression of CD133 and Nanog induced by treatment with ZSTK474, detected by RT-PCR. * P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Prolonged inhibition of class I PI3K promotes liver cancer stem cell expansion by augmenting SGK3/GSK-3β/β-catenin signalling

    doi: 10.1186/s13046-018-0801-8

    Figure Lengend Snippet: The inhibitor of hVps34 can block SGK3 activity and suppress liver CSC expansion after prolonged treatment of HCC cells with PI3K inhibitors. a Huh7 and MHCC97H cells were treated with VPS34-IN1 for 24 h, and cell lysates were subjected to western blot analysis with the indicated antibodies. b Spheroid formation assay of MHCC97H cells treated with VPS34-IN1 (top), the statistical results of the tumour spheroids ( > 50 μm) were calculated based on 3 independent experiments (bottom). Scale bars, 200 μm. c Immunofluorescence images of Nanog (red) in MHCC7H cells after treatment with VPS34-IN1 or dimethyl sulphoxide DMSO for 24 h. Nuclei were stained with DAPI (blue). Scale bars, 10 μm. d The proportion of CD133+ cells in Huh7 cells was evaluated by flow cytometric assay. e Treatment of MHCC97H cells with VPS34-IN1 for 24 h inhibited the overexpression of CD133 and Nanog induced by treatment with ZSTK474, detected by RT-PCR. * P

    Article Snippet: Sections were deparaffinized and stained with anti-CD133 (1:500; ab222782; Abcam) at 4 °C overnight, and incubated with the secondary antibody for 1 h at 37 °C.

    Techniques: Blocking Assay, Activity Assay, Western Blot, Tube Formation Assay, Immunofluorescence, Staining, Flow Cytometry, Over Expression, Reverse Transcription Polymerase Chain Reaction

    SGK3 promotes liver CSCs through β-catenin accumulation by GSK3β inactivation. a Levels of β-catenin and active GSK3β (phosphorylated at Ser9)/total GSK3β compared between MHCC97H spheroids and attached cells detected by western blot. β-actin was used as a loading control. b Expression of β-catenin, active GSK3β (phosphorylated at Ser9)/ total GSK3β and SGK3 in Huh7 and MHCC-97H cells stably overexpressing SGK3 or NC detected by western blot. β-actin was used as a loading control. c Expression of β-catenin, active GSK3β (phosphorylated at Ser9)/total GSK3β and SGK3 in Huh7 SGK3 shRNA cells or control cells was detected by western blot. d Treatment of MHCC97H cells with AR-A014418 for 24 h inhibited the expression of CD133, detected by RT-PCR. e MHCC97H-SGK3 or MHCC-97H-NC were treated with 20 μM of GSK3β inhibitor AR-A014418 and subjected to spheroid formation assay (left). The statistical results of the tumour spheroids ( > 50 μm) were calculated based on 3 independent experiments (right). Scale bars, 200 μm. All experiments were performed in triplicate, and the results are shown as the mean ± standard deviation. * P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Prolonged inhibition of class I PI3K promotes liver cancer stem cell expansion by augmenting SGK3/GSK-3β/β-catenin signalling

    doi: 10.1186/s13046-018-0801-8

    Figure Lengend Snippet: SGK3 promotes liver CSCs through β-catenin accumulation by GSK3β inactivation. a Levels of β-catenin and active GSK3β (phosphorylated at Ser9)/total GSK3β compared between MHCC97H spheroids and attached cells detected by western blot. β-actin was used as a loading control. b Expression of β-catenin, active GSK3β (phosphorylated at Ser9)/ total GSK3β and SGK3 in Huh7 and MHCC-97H cells stably overexpressing SGK3 or NC detected by western blot. β-actin was used as a loading control. c Expression of β-catenin, active GSK3β (phosphorylated at Ser9)/total GSK3β and SGK3 in Huh7 SGK3 shRNA cells or control cells was detected by western blot. d Treatment of MHCC97H cells with AR-A014418 for 24 h inhibited the expression of CD133, detected by RT-PCR. e MHCC97H-SGK3 or MHCC-97H-NC were treated with 20 μM of GSK3β inhibitor AR-A014418 and subjected to spheroid formation assay (left). The statistical results of the tumour spheroids ( > 50 μm) were calculated based on 3 independent experiments (right). Scale bars, 200 μm. All experiments were performed in triplicate, and the results are shown as the mean ± standard deviation. * P

    Article Snippet: Sections were deparaffinized and stained with anti-CD133 (1:500; ab222782; Abcam) at 4 °C overnight, and incubated with the secondary antibody for 1 h at 37 °C.

    Techniques: Western Blot, Expressing, Stable Transfection, shRNA, Reverse Transcription Polymerase Chain Reaction, Tube Formation Assay, Standard Deviation

    SGK3 is preferentially activated in liver CSCs. a MHCC-97H cells were cultured in monolayer or ultra-low attachment conditions. The mRNA expression of liver CSC-related genes and SGK3 in spheroids and attached cells was compared by RT-PCR. b Western blot analysis for levels of CD133, active Akt (phosphorylated at Ser473)/total Akt and active SGK3 (phosphorylated at Thr320)/total SGK3 between spheroids and attached cells. c Flow cytometry analysis of CD133+ cell distribution in CD133- and CD133+ cells isolated using CD133 MicroBead Kit. d Representative images of CD133+ and CD133- cells sorted from MHCC97H HCC cells cultured in serum-free culture medium after 7 days. Scale bars, 100 μm. e CD133+/CD133- cells were subcutaneously injected into 6-week-old female athymic nude mice, and tumourigenicity was evaluated 5 weeks post-inoculation. f Levels of CD133, active Akt (phosphorylated at Ser473)/total Akt and active SGK3 (phosphorylated at Thr320)/total SGK3 were compared by western blot analysis between CD133+ and CD133- cells. β-actin was used as a loading control. All experiments were performed in triplicate. * P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Prolonged inhibition of class I PI3K promotes liver cancer stem cell expansion by augmenting SGK3/GSK-3β/β-catenin signalling

    doi: 10.1186/s13046-018-0801-8

    Figure Lengend Snippet: SGK3 is preferentially activated in liver CSCs. a MHCC-97H cells were cultured in monolayer or ultra-low attachment conditions. The mRNA expression of liver CSC-related genes and SGK3 in spheroids and attached cells was compared by RT-PCR. b Western blot analysis for levels of CD133, active Akt (phosphorylated at Ser473)/total Akt and active SGK3 (phosphorylated at Thr320)/total SGK3 between spheroids and attached cells. c Flow cytometry analysis of CD133+ cell distribution in CD133- and CD133+ cells isolated using CD133 MicroBead Kit. d Representative images of CD133+ and CD133- cells sorted from MHCC97H HCC cells cultured in serum-free culture medium after 7 days. Scale bars, 100 μm. e CD133+/CD133- cells were subcutaneously injected into 6-week-old female athymic nude mice, and tumourigenicity was evaluated 5 weeks post-inoculation. f Levels of CD133, active Akt (phosphorylated at Ser473)/total Akt and active SGK3 (phosphorylated at Thr320)/total SGK3 were compared by western blot analysis between CD133+ and CD133- cells. β-actin was used as a loading control. All experiments were performed in triplicate. * P

    Article Snippet: Sections were deparaffinized and stained with anti-CD133 (1:500; ab222782; Abcam) at 4 °C overnight, and incubated with the secondary antibody for 1 h at 37 °C.

    Techniques: Cell Culture, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Flow Cytometry, Cytometry, Isolation, Injection, Mouse Assay

    SGK3 enhances the expansion of liver CSCs. a, b Relative mRNA expression of liver CSC-related markers CD133, CD90, Nanog, Oct4, Bmi-1 and Sox2 in Huh7 and MHCC-97H cells stably overexpressing SGK3 or NC. β-actin was used as an loading control. c Expression of CD133, Nanog and SGK3 in Huh7 and MHCC-97H cells stably overexpressing SGK3 or NC detected by western blot. β-actin was used as a loading control. d Spheroid formation assay of MHCC97H-SGK3 or MHCC97H-NC cells (top). Scale bars, 200 μm. The statistical results of the tumour spherosis ( > 50 μm) were calculated based on 3 independent experiments (bottom). All experiments were performed in triplicate, and the results are shown as mean ± standard deviation. * P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Prolonged inhibition of class I PI3K promotes liver cancer stem cell expansion by augmenting SGK3/GSK-3β/β-catenin signalling

    doi: 10.1186/s13046-018-0801-8

    Figure Lengend Snippet: SGK3 enhances the expansion of liver CSCs. a, b Relative mRNA expression of liver CSC-related markers CD133, CD90, Nanog, Oct4, Bmi-1 and Sox2 in Huh7 and MHCC-97H cells stably overexpressing SGK3 or NC. β-actin was used as an loading control. c Expression of CD133, Nanog and SGK3 in Huh7 and MHCC-97H cells stably overexpressing SGK3 or NC detected by western blot. β-actin was used as a loading control. d Spheroid formation assay of MHCC97H-SGK3 or MHCC97H-NC cells (top). Scale bars, 200 μm. The statistical results of the tumour spherosis ( > 50 μm) were calculated based on 3 independent experiments (bottom). All experiments were performed in triplicate, and the results are shown as mean ± standard deviation. * P

    Article Snippet: Sections were deparaffinized and stained with anti-CD133 (1:500; ab222782; Abcam) at 4 °C overnight, and incubated with the secondary antibody for 1 h at 37 °C.

    Techniques: Expressing, Stable Transfection, Western Blot, Tube Formation Assay, Standard Deviation

    Inhibition of SGK3 attenuates liver CSC expansion. a, b Huh7-transfected SGK3 shRNA1 or MHCC97H-transfected SGK3 shRNA1 and their control cells were collected and subjected to real-time PCR. β-actin was used as a loading control. c Expression of Nanog, CD133, and SGK3 was detected by western blot in Huh7 shRNA1 or MHCC97H shRNA1 and their control cells. β-actin was used as a loading control. d Spheroid formation assay of SGK3 knockdown MHCC97H cells (MHCC97H-SGK3-shRNA1) or control cells (MHCC97H-SGK3-control) (up). Scale bars, 200 μm. The statistical results of the tumour spheroids ( > 50 μm) were calculated based on 3 independent experiments (down). e The proportion of CD133+ cells in SGK3 knockdown MHCC97H cells or control cells was evaluated by flow cytometric assay. All experiments were performed in triplicate, and the results are shown as mean ± standard deviation. * P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Prolonged inhibition of class I PI3K promotes liver cancer stem cell expansion by augmenting SGK3/GSK-3β/β-catenin signalling

    doi: 10.1186/s13046-018-0801-8

    Figure Lengend Snippet: Inhibition of SGK3 attenuates liver CSC expansion. a, b Huh7-transfected SGK3 shRNA1 or MHCC97H-transfected SGK3 shRNA1 and their control cells were collected and subjected to real-time PCR. β-actin was used as a loading control. c Expression of Nanog, CD133, and SGK3 was detected by western blot in Huh7 shRNA1 or MHCC97H shRNA1 and their control cells. β-actin was used as a loading control. d Spheroid formation assay of SGK3 knockdown MHCC97H cells (MHCC97H-SGK3-shRNA1) or control cells (MHCC97H-SGK3-control) (up). Scale bars, 200 μm. The statistical results of the tumour spheroids ( > 50 μm) were calculated based on 3 independent experiments (down). e The proportion of CD133+ cells in SGK3 knockdown MHCC97H cells or control cells was evaluated by flow cytometric assay. All experiments were performed in triplicate, and the results are shown as mean ± standard deviation. * P

    Article Snippet: Sections were deparaffinized and stained with anti-CD133 (1:500; ab222782; Abcam) at 4 °C overnight, and incubated with the secondary antibody for 1 h at 37 °C.

    Techniques: Inhibition, Transfection, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Tube Formation Assay, Flow Cytometry, Standard Deviation