mouse prb Search Results


mouse rb 1  (OriGene)
90
OriGene mouse rb 1
a. Glutamine uptake in WT, <t>Rb-1</t> −/− and TKO cells was assayed by 14 C-glutamine labeling. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.005 . b. Glutamine uptake was determined in Rb-1 −/− and TKO cells following reconstitution of Rb-1 expression. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.01 . c. mRNA expression of ASCT2 and SLC38A5 between WT and TKO MEFs. For the ASCT2 and SLC38A5 comparisons, data are represented as relative mRNA with WT levels set to 1. Protein expression of ASCT2 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . d. For GLS, mRNA levels are relative to expression of WT GLS1 set to 1. For mRNA, shown is mean ± s.d. from three independent RNA preparations. Protein expression of GLS1 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . e. Secreted ammonia levels were determined in the medium from WT and TKO cells. Data are represented as nmol / min / mg protein, and shown are mean ± s.d. from triplicate measurements from two independent experiments. * p < 0.005 . f. Incorporation of glutamine carbon into glutamate was determined by stable isotope labeling of WT and TKO cells with [ 13 C 5 , 15 N 2 ]-glutamine, as described in the methods. Shown are representative 2-D NMR spectra of peaks representative of glutamate. The boxes connect the 13 C satellite peaks of glutamate and the central peak denoted 12 C is from glutamate that contains no 13 C.
Mouse Rb 1, supplied by OriGene, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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control mouse monoclonal antibody, 13d210, prb  (Cold Spring Harbor Laboratory Meetings)
90
Cold Spring Harbor Laboratory Meetings control mouse monoclonal antibody, 13d210, prb
a. Glutamine uptake in WT, <t>Rb-1</t> −/− and TKO cells was assayed by 14 C-glutamine labeling. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.005 . b. Glutamine uptake was determined in Rb-1 −/− and TKO cells following reconstitution of Rb-1 expression. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.01 . c. mRNA expression of ASCT2 and SLC38A5 between WT and TKO MEFs. For the ASCT2 and SLC38A5 comparisons, data are represented as relative mRNA with WT levels set to 1. Protein expression of ASCT2 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . d. For GLS, mRNA levels are relative to expression of WT GLS1 set to 1. For mRNA, shown is mean ± s.d. from three independent RNA preparations. Protein expression of GLS1 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . e. Secreted ammonia levels were determined in the medium from WT and TKO cells. Data are represented as nmol / min / mg protein, and shown are mean ± s.d. from triplicate measurements from two independent experiments. * p < 0.005 . f. Incorporation of glutamine carbon into glutamate was determined by stable isotope labeling of WT and TKO cells with [ 13 C 5 , 15 N 2 ]-glutamine, as described in the methods. Shown are representative 2-D NMR spectra of peaks representative of glutamate. The boxes connect the 13 C satellite peaks of glutamate and the central peak denoted 12 C is from glutamate that contains no 13 C.
Control Mouse Monoclonal Antibody, 13d210, Prb, supplied by Cold Spring Harbor Laboratory Meetings, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse monoclonal antibodies retinoblastoma protein (prb  (Becton Dickinson)
90
Becton Dickinson mouse monoclonal antibodies retinoblastoma protein (prb
a. Glutamine uptake in WT, <t>Rb-1</t> −/− and TKO cells was assayed by 14 C-glutamine labeling. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.005 . b. Glutamine uptake was determined in Rb-1 −/− and TKO cells following reconstitution of Rb-1 expression. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.01 . c. mRNA expression of ASCT2 and SLC38A5 between WT and TKO MEFs. For the ASCT2 and SLC38A5 comparisons, data are represented as relative mRNA with WT levels set to 1. Protein expression of ASCT2 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . d. For GLS, mRNA levels are relative to expression of WT GLS1 set to 1. For mRNA, shown is mean ± s.d. from three independent RNA preparations. Protein expression of GLS1 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . e. Secreted ammonia levels were determined in the medium from WT and TKO cells. Data are represented as nmol / min / mg protein, and shown are mean ± s.d. from triplicate measurements from two independent experiments. * p < 0.005 . f. Incorporation of glutamine carbon into glutamate was determined by stable isotope labeling of WT and TKO cells with [ 13 C 5 , 15 N 2 ]-glutamine, as described in the methods. Shown are representative 2-D NMR spectra of peaks representative of glutamate. The boxes connect the 13 C satellite peaks of glutamate and the central peak denoted 12 C is from glutamate that contains no 13 C.
Mouse Monoclonal Antibodies Retinoblastoma Protein (Prb, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti-mouse prb  (Oncogene Science Inc)
90
Oncogene Science Inc anti-mouse prb
a. Glutamine uptake in WT, <t>Rb-1</t> −/− and TKO cells was assayed by 14 C-glutamine labeling. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.005 . b. Glutamine uptake was determined in Rb-1 −/− and TKO cells following reconstitution of Rb-1 expression. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.01 . c. mRNA expression of ASCT2 and SLC38A5 between WT and TKO MEFs. For the ASCT2 and SLC38A5 comparisons, data are represented as relative mRNA with WT levels set to 1. Protein expression of ASCT2 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . d. For GLS, mRNA levels are relative to expression of WT GLS1 set to 1. For mRNA, shown is mean ± s.d. from three independent RNA preparations. Protein expression of GLS1 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . e. Secreted ammonia levels were determined in the medium from WT and TKO cells. Data are represented as nmol / min / mg protein, and shown are mean ± s.d. from triplicate measurements from two independent experiments. * p < 0.005 . f. Incorporation of glutamine carbon into glutamate was determined by stable isotope labeling of WT and TKO cells with [ 13 C 5 , 15 N 2 ]-glutamine, as described in the methods. Shown are representative 2-D NMR spectra of peaks representative of glutamate. The boxes connect the 13 C satellite peaks of glutamate and the central peak denoted 12 C is from glutamate that contains no 13 C.
Anti Mouse Prb, supplied by Oncogene Science Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse filaggrin (cat. no. prb-417p; dilution 1:5000)  (Nordic BioSite)
90
Nordic BioSite mouse filaggrin (cat. no. prb-417p; dilution 1:5000)
a. Glutamine uptake in WT, <t>Rb-1</t> −/− and TKO cells was assayed by 14 C-glutamine labeling. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.005 . b. Glutamine uptake was determined in Rb-1 −/− and TKO cells following reconstitution of Rb-1 expression. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.01 . c. mRNA expression of ASCT2 and SLC38A5 between WT and TKO MEFs. For the ASCT2 and SLC38A5 comparisons, data are represented as relative mRNA with WT levels set to 1. Protein expression of ASCT2 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . d. For GLS, mRNA levels are relative to expression of WT GLS1 set to 1. For mRNA, shown is mean ± s.d. from three independent RNA preparations. Protein expression of GLS1 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . e. Secreted ammonia levels were determined in the medium from WT and TKO cells. Data are represented as nmol / min / mg protein, and shown are mean ± s.d. from triplicate measurements from two independent experiments. * p < 0.005 . f. Incorporation of glutamine carbon into glutamate was determined by stable isotope labeling of WT and TKO cells with [ 13 C 5 , 15 N 2 ]-glutamine, as described in the methods. Shown are representative 2-D NMR spectra of peaks representative of glutamate. The boxes connect the 13 C satellite peaks of glutamate and the central peak denoted 12 C is from glutamate that contains no 13 C.
Mouse Filaggrin (Cat. No. Prb 417p; Dilution 1:5000), supplied by Nordic BioSite, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse prb  (Chester Beatty)
90
Chester Beatty mouse prb
a. Glutamine uptake in WT, <t>Rb-1</t> −/− and TKO cells was assayed by 14 C-glutamine labeling. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.005 . b. Glutamine uptake was determined in Rb-1 −/− and TKO cells following reconstitution of Rb-1 expression. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.01 . c. mRNA expression of ASCT2 and SLC38A5 between WT and TKO MEFs. For the ASCT2 and SLC38A5 comparisons, data are represented as relative mRNA with WT levels set to 1. Protein expression of ASCT2 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . d. For GLS, mRNA levels are relative to expression of WT GLS1 set to 1. For mRNA, shown is mean ± s.d. from three independent RNA preparations. Protein expression of GLS1 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . e. Secreted ammonia levels were determined in the medium from WT and TKO cells. Data are represented as nmol / min / mg protein, and shown are mean ± s.d. from triplicate measurements from two independent experiments. * p < 0.005 . f. Incorporation of glutamine carbon into glutamate was determined by stable isotope labeling of WT and TKO cells with [ 13 C 5 , 15 N 2 ]-glutamine, as described in the methods. Shown are representative 2-D NMR spectra of peaks representative of glutamate. The boxes connect the 13 C satellite peaks of glutamate and the central peak denoted 12 C is from glutamate that contains no 13 C.
Mouse Prb, supplied by Chester Beatty, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse anti-prb  (Becton Dickinson)
90
Becton Dickinson mouse anti-prb
a. Glutamine uptake in WT, <t>Rb-1</t> −/− and TKO cells was assayed by 14 C-glutamine labeling. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.005 . b. Glutamine uptake was determined in Rb-1 −/− and TKO cells following reconstitution of Rb-1 expression. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.01 . c. mRNA expression of ASCT2 and SLC38A5 between WT and TKO MEFs. For the ASCT2 and SLC38A5 comparisons, data are represented as relative mRNA with WT levels set to 1. Protein expression of ASCT2 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . d. For GLS, mRNA levels are relative to expression of WT GLS1 set to 1. For mRNA, shown is mean ± s.d. from three independent RNA preparations. Protein expression of GLS1 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . e. Secreted ammonia levels were determined in the medium from WT and TKO cells. Data are represented as nmol / min / mg protein, and shown are mean ± s.d. from triplicate measurements from two independent experiments. * p < 0.005 . f. Incorporation of glutamine carbon into glutamate was determined by stable isotope labeling of WT and TKO cells with [ 13 C 5 , 15 N 2 ]-glutamine, as described in the methods. Shown are representative 2-D NMR spectra of peaks representative of glutamate. The boxes connect the 13 C satellite peaks of glutamate and the central peak denoted 12 C is from glutamate that contains no 13 C.
Mouse Anti Prb, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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prb (mouse  (Becton Dickinson)
90
Becton Dickinson prb (mouse
Primary and secondary antibodies used for immunofluorescence.
Prb (Mouse, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse monoclonal antibody human retinoblastoma protein (prb  (Becton Dickinson)
90
Becton Dickinson mouse monoclonal antibody human retinoblastoma protein (prb
Primary and secondary antibodies used for immunofluorescence.
Mouse Monoclonal Antibody Human Retinoblastoma Protein (Prb, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse monoclonal anti prb 554136  (Becton Dickinson)
90
Becton Dickinson mouse monoclonal anti prb 554136
<t>pRb</t> depletion causes centrosome amplification, aneuploidy and altered gene expression . A) RT-PCR analysis for RB expression in IMR90 cells transfected with siRNA-luciferase (siLuc), siRNA targeting RB (siRB) and after RB re-expression (release). B) The histogram (top panel) summarizes the percentage of cells with 1, 2 or >2 centrosomes in control and pRb-depleted IMR90 cells. Presence of supernumerary centrosomes in interphase cells (bottom panel: b) and abnormal mitotic spindles (bottom panel: c, arrowheads) in IMR90 cells transfected with siRNA targeting RB in comparison with control cells (bottom panel: a) as revealed by immunofluorescence for γ-tubulin (green) or β-tubulin (green) respectively, nuclei were stained with DAPI (blue). C) Bar graphs showing the percentage of cells with normal (46 chromosomes), hypodiploid (<46 chromosomes) and hyperdiploid (>46 chromosomes) metaphases after RB silencing and in control cells, untransfected or transfected with siRNA-luciferase (siLuc). D) Expression levels of genes involved in mitosis progression by Real time RT-PCR. The x-axis indicates genes and the y-axis the relative quantification in IMR90 cells untransfected (wt), transfected with siRNA-luciferase (siLuc) and siRNA targeting RB (siRB). E) Western blot analysis in pRb silenced IMR90 cells showing lack of pRb, increase of <t>Aurora-A,</t> <t>Plk1</t> and decrease of Mad2 protein at 72 hours. β-actin is used a loading control.
Mouse Monoclonal Anti Prb 554136, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse anti-prb pser612 clone 4e4  (Cyclex Inc)
90
Cyclex Inc mouse anti-prb pser612 clone 4e4
<t>pRb</t> depletion causes centrosome amplification, aneuploidy and altered gene expression . A) RT-PCR analysis for RB expression in IMR90 cells transfected with siRNA-luciferase (siLuc), siRNA targeting RB (siRB) and after RB re-expression (release). B) The histogram (top panel) summarizes the percentage of cells with 1, 2 or >2 centrosomes in control and pRb-depleted IMR90 cells. Presence of supernumerary centrosomes in interphase cells (bottom panel: b) and abnormal mitotic spindles (bottom panel: c, arrowheads) in IMR90 cells transfected with siRNA targeting RB in comparison with control cells (bottom panel: a) as revealed by immunofluorescence for γ-tubulin (green) or β-tubulin (green) respectively, nuclei were stained with DAPI (blue). C) Bar graphs showing the percentage of cells with normal (46 chromosomes), hypodiploid (<46 chromosomes) and hyperdiploid (>46 chromosomes) metaphases after RB silencing and in control cells, untransfected or transfected with siRNA-luciferase (siLuc). D) Expression levels of genes involved in mitosis progression by Real time RT-PCR. The x-axis indicates genes and the y-axis the relative quantification in IMR90 cells untransfected (wt), transfected with siRNA-luciferase (siLuc) and siRNA targeting RB (siRB). E) Western blot analysis in pRb silenced IMR90 cells showing lack of pRb, increase of <t>Aurora-A,</t> <t>Plk1</t> and decrease of Mad2 protein at 72 hours. β-actin is used a loading control.
Mouse Anti Prb Pser612 Clone 4e4, supplied by Cyclex Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti-mouse antibodies prb 145p  (HiSS Diagnostics)
90
HiSS Diagnostics rabbit anti-mouse antibodies prb 145p
<t>pRb</t> depletion causes centrosome amplification, aneuploidy and altered gene expression . A) RT-PCR analysis for RB expression in IMR90 cells transfected with siRNA-luciferase (siLuc), siRNA targeting RB (siRB) and after RB re-expression (release). B) The histogram (top panel) summarizes the percentage of cells with 1, 2 or >2 centrosomes in control and pRb-depleted IMR90 cells. Presence of supernumerary centrosomes in interphase cells (bottom panel: b) and abnormal mitotic spindles (bottom panel: c, arrowheads) in IMR90 cells transfected with siRNA targeting RB in comparison with control cells (bottom panel: a) as revealed by immunofluorescence for γ-tubulin (green) or β-tubulin (green) respectively, nuclei were stained with DAPI (blue). C) Bar graphs showing the percentage of cells with normal (46 chromosomes), hypodiploid (<46 chromosomes) and hyperdiploid (>46 chromosomes) metaphases after RB silencing and in control cells, untransfected or transfected with siRNA-luciferase (siLuc). D) Expression levels of genes involved in mitosis progression by Real time RT-PCR. The x-axis indicates genes and the y-axis the relative quantification in IMR90 cells untransfected (wt), transfected with siRNA-luciferase (siLuc) and siRNA targeting RB (siRB). E) Western blot analysis in pRb silenced IMR90 cells showing lack of pRb, increase of <t>Aurora-A,</t> <t>Plk1</t> and decrease of Mad2 protein at 72 hours. β-actin is used a loading control.
Rabbit Anti Mouse Antibodies Prb 145p, supplied by HiSS Diagnostics, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


a. Glutamine uptake in WT, Rb-1 −/− and TKO cells was assayed by 14 C-glutamine labeling. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.005 . b. Glutamine uptake was determined in Rb-1 −/− and TKO cells following reconstitution of Rb-1 expression. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.01 . c. mRNA expression of ASCT2 and SLC38A5 between WT and TKO MEFs. For the ASCT2 and SLC38A5 comparisons, data are represented as relative mRNA with WT levels set to 1. Protein expression of ASCT2 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . d. For GLS, mRNA levels are relative to expression of WT GLS1 set to 1. For mRNA, shown is mean ± s.d. from three independent RNA preparations. Protein expression of GLS1 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . e. Secreted ammonia levels were determined in the medium from WT and TKO cells. Data are represented as nmol / min / mg protein, and shown are mean ± s.d. from triplicate measurements from two independent experiments. * p < 0.005 . f. Incorporation of glutamine carbon into glutamate was determined by stable isotope labeling of WT and TKO cells with [ 13 C 5 , 15 N 2 ]-glutamine, as described in the methods. Shown are representative 2-D NMR spectra of peaks representative of glutamate. The boxes connect the 13 C satellite peaks of glutamate and the central peak denoted 12 C is from glutamate that contains no 13 C.

Journal: Oncogene

Article Title: Control of Glutamine Metabolism By the Tumor Suppressor Rb

doi: 10.1038/onc.2012.635

Figure Lengend Snippet: a. Glutamine uptake in WT, Rb-1 −/− and TKO cells was assayed by 14 C-glutamine labeling. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.005 . b. Glutamine uptake was determined in Rb-1 −/− and TKO cells following reconstitution of Rb-1 expression. Data are represented as nmol / min / mg protein. Mean ± s.d. from triplicate measurements from three independent cell preparations. * p < 0.01 . c. mRNA expression of ASCT2 and SLC38A5 between WT and TKO MEFs. For the ASCT2 and SLC38A5 comparisons, data are represented as relative mRNA with WT levels set to 1. Protein expression of ASCT2 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . d. For GLS, mRNA levels are relative to expression of WT GLS1 set to 1. For mRNA, shown is mean ± s.d. from three independent RNA preparations. Protein expression of GLS1 between WT and TKO MEFs is represented as relative protein expression with WT levels set to 1. For protein expression, shown is a representative image from three separate experiments. * p < 0.01 . e. Secreted ammonia levels were determined in the medium from WT and TKO cells. Data are represented as nmol / min / mg protein, and shown are mean ± s.d. from triplicate measurements from two independent experiments. * p < 0.005 . f. Incorporation of glutamine carbon into glutamate was determined by stable isotope labeling of WT and TKO cells with [ 13 C 5 , 15 N 2 ]-glutamine, as described in the methods. Shown are representative 2-D NMR spectra of peaks representative of glutamate. The boxes connect the 13 C satellite peaks of glutamate and the central peak denoted 12 C is from glutamate that contains no 13 C.

Article Snippet: Cells were transfected with a plasmid encoding mouse Rb-1 (Origene: MG227287) using JetPrime reagent following manufacturer’s protocol (Polyplus). siRNA transfections were performed as previously described ( ) and the following siRNAs were used: E2F-1: (Ambion: #s201266); E2F-2: (Ambion: #s109846); E2F-3: (Ambion: #s65238).

Techniques: Labeling, Expressing

Primary and secondary antibodies used for immunofluorescence.

Journal: PLoS ONE

Article Title: Polyomavirus-Associated Trichodysplasia Spinulosa Involves Hyperproliferation, pRB Phosphorylation and Upregulation of p16 and p21

doi: 10.1371/journal.pone.0108947

Figure Lengend Snippet: Primary and secondary antibodies used for immunofluorescence.

Article Snippet: pRB (mouse) , G3-245 , 1∶250 , BD Biosciences, USA.

Techniques: Immunofluorescence

pRb depletion causes centrosome amplification, aneuploidy and altered gene expression . A) RT-PCR analysis for RB expression in IMR90 cells transfected with siRNA-luciferase (siLuc), siRNA targeting RB (siRB) and after RB re-expression (release). B) The histogram (top panel) summarizes the percentage of cells with 1, 2 or >2 centrosomes in control and pRb-depleted IMR90 cells. Presence of supernumerary centrosomes in interphase cells (bottom panel: b) and abnormal mitotic spindles (bottom panel: c, arrowheads) in IMR90 cells transfected with siRNA targeting RB in comparison with control cells (bottom panel: a) as revealed by immunofluorescence for γ-tubulin (green) or β-tubulin (green) respectively, nuclei were stained with DAPI (blue). C) Bar graphs showing the percentage of cells with normal (46 chromosomes), hypodiploid (<46 chromosomes) and hyperdiploid (>46 chromosomes) metaphases after RB silencing and in control cells, untransfected or transfected with siRNA-luciferase (siLuc). D) Expression levels of genes involved in mitosis progression by Real time RT-PCR. The x-axis indicates genes and the y-axis the relative quantification in IMR90 cells untransfected (wt), transfected with siRNA-luciferase (siLuc) and siRNA targeting RB (siRB). E) Western blot analysis in pRb silenced IMR90 cells showing lack of pRb, increase of Aurora-A, Plk1 and decrease of Mad2 protein at 72 hours. β-actin is used a loading control.

Journal: BMC Cell Biology

Article Title: RNAi mediated acute depletion of Retinoblastoma protein (pRb) promotes aneuploidy in human primary cells via micronuclei formation

doi: 10.1186/1471-2121-10-79

Figure Lengend Snippet: pRb depletion causes centrosome amplification, aneuploidy and altered gene expression . A) RT-PCR analysis for RB expression in IMR90 cells transfected with siRNA-luciferase (siLuc), siRNA targeting RB (siRB) and after RB re-expression (release). B) The histogram (top panel) summarizes the percentage of cells with 1, 2 or >2 centrosomes in control and pRb-depleted IMR90 cells. Presence of supernumerary centrosomes in interphase cells (bottom panel: b) and abnormal mitotic spindles (bottom panel: c, arrowheads) in IMR90 cells transfected with siRNA targeting RB in comparison with control cells (bottom panel: a) as revealed by immunofluorescence for γ-tubulin (green) or β-tubulin (green) respectively, nuclei were stained with DAPI (blue). C) Bar graphs showing the percentage of cells with normal (46 chromosomes), hypodiploid (<46 chromosomes) and hyperdiploid (>46 chromosomes) metaphases after RB silencing and in control cells, untransfected or transfected with siRNA-luciferase (siLuc). D) Expression levels of genes involved in mitosis progression by Real time RT-PCR. The x-axis indicates genes and the y-axis the relative quantification in IMR90 cells untransfected (wt), transfected with siRNA-luciferase (siLuc) and siRNA targeting RB (siRB). E) Western blot analysis in pRb silenced IMR90 cells showing lack of pRb, increase of Aurora-A, Plk1 and decrease of Mad2 protein at 72 hours. β-actin is used a loading control.

Article Snippet: Western blot was probed with mouse monoclonal anti pRB (554136, Becton Dickinson), anti Plk1 (sc-17783 F-8, Santa Cruz) or anti Cyclin-E (SC-247he12 Santa Cruz Biotechnology, Inc.) antibodies.

Techniques: Amplification, Gene Expression, Reverse Transcription Polymerase Chain Reaction, Expressing, Transfection, Luciferase, Control, Comparison, Immunofluorescence, Staining, Quantitative RT-PCR, Quantitative Proteomics, Western Blot

Evaluation of cell growth at different times after RB silencing . A) Decreased proliferation rates starting 48 h up to 7 days from transfection in RB/PLK1 and RB/AURORA-A co-depleted cells (double knock-down) in comparison with both untransfected and pRb-depleted cells. B) Western blot analysis showing increased Cyclin-E protein levels in pRb-depleted cells as well as in cells with knock-down of both RB and AURORA-A or PLK1 , and increased Cyclin-A protein levels in RB/AURORA-A co-depleted cells. Cells with knock-down of both RB and AURORA-A or PLK1 but not with RB knock-down showed increased p21 Cip1 levels. β-tubulin was used as a loading control of protein extracts.

Journal: BMC Cell Biology

Article Title: RNAi mediated acute depletion of Retinoblastoma protein (pRb) promotes aneuploidy in human primary cells via micronuclei formation

doi: 10.1186/1471-2121-10-79

Figure Lengend Snippet: Evaluation of cell growth at different times after RB silencing . A) Decreased proliferation rates starting 48 h up to 7 days from transfection in RB/PLK1 and RB/AURORA-A co-depleted cells (double knock-down) in comparison with both untransfected and pRb-depleted cells. B) Western blot analysis showing increased Cyclin-E protein levels in pRb-depleted cells as well as in cells with knock-down of both RB and AURORA-A or PLK1 , and increased Cyclin-A protein levels in RB/AURORA-A co-depleted cells. Cells with knock-down of both RB and AURORA-A or PLK1 but not with RB knock-down showed increased p21 Cip1 levels. β-tubulin was used as a loading control of protein extracts.

Article Snippet: Western blot was probed with mouse monoclonal anti pRB (554136, Becton Dickinson), anti Plk1 (sc-17783 F-8, Santa Cruz) or anti Cyclin-E (SC-247he12 Santa Cruz Biotechnology, Inc.) antibodies.

Techniques: Transfection, Knockdown, Comparison, Western Blot, Control

Micronuclei generation after pRb acute loss in primary human fibroblasts . A) Bar graphs showing micronuclei percentages scored in RB transcriptionally silenced IMR90 cells (siRB) and in IMR90 cells with knock-down of both RB and AURORA-A or PLK1 compared with untransfected cells (untr.) or cells transfected with siRNA-luciferase (siLuc). B) Immunofluorescence microscopy analysis detecting the presence of centromeres (green spots) in IMR90 wild-type cells (b), in micronuclei generated following RB transcriptional silencing (c), and after knock-down of both RB and PLK1 (d) or after knock-down of both RB and AURORA-A (e). In (a) are shown lagging chromosomes (arrow) observed during metaphases in RB transcriptionally silenced cells. Nuclei were stained with DAPI (blue).

Journal: BMC Cell Biology

Article Title: RNAi mediated acute depletion of Retinoblastoma protein (pRb) promotes aneuploidy in human primary cells via micronuclei formation

doi: 10.1186/1471-2121-10-79

Figure Lengend Snippet: Micronuclei generation after pRb acute loss in primary human fibroblasts . A) Bar graphs showing micronuclei percentages scored in RB transcriptionally silenced IMR90 cells (siRB) and in IMR90 cells with knock-down of both RB and AURORA-A or PLK1 compared with untransfected cells (untr.) or cells transfected with siRNA-luciferase (siLuc). B) Immunofluorescence microscopy analysis detecting the presence of centromeres (green spots) in IMR90 wild-type cells (b), in micronuclei generated following RB transcriptional silencing (c), and after knock-down of both RB and PLK1 (d) or after knock-down of both RB and AURORA-A (e). In (a) are shown lagging chromosomes (arrow) observed during metaphases in RB transcriptionally silenced cells. Nuclei were stained with DAPI (blue).

Article Snippet: Western blot was probed with mouse monoclonal anti pRB (554136, Becton Dickinson), anti Plk1 (sc-17783 F-8, Santa Cruz) or anti Cyclin-E (SC-247he12 Santa Cruz Biotechnology, Inc.) antibodies.

Techniques: Knockdown, Transfection, Luciferase, Immunofluorescence, Microscopy, Generated, Staining