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osrc2 cells  (ATCC)


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    ATCC osrc2 cells
    Osrc2 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 2388 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    cell lines osrc2  (Procell Inc)
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    pVHL mediates the ubiquitination and degradation of CYP1B1. A and B Assessment of CYP1B1 mRNA and protein levels in ACHN cells with stable VHL knockdown and 786O and <t>OSRC2</t> cells with stable VHL overexpression via western blotting and RT‒qPCR. C Coimmunoprecipitation of ACHN cell whole-cell lysates with CYP1B1, pVHL or IgG antibodies, followed by western blotting with the respective antibodies. D 293T and 786O cells were transfected with Flag-CYP1B1 and His-pVHL plasmids. Coimmunoprecipitation with CYP1B1, pVHL or IgG antibodies was followed by western blotting with the indicated antibodies. E GST affinity-isolation assays with purified tagged pVHL and CYP1B1 proteins, followed by western blotting analysis. F Cotransfection of 293T and ACHN cells with VHL and CYP1B1 overexpression plasmids and subsequent immunofluorescence staining with anti-pVHL and anti-CYP1B1 antibodies. Scale bar: 10 μm. G Protein synthesis inhibition via CHX (10 μM) in 786O cells with VHL overexpression and ACHN cells with VHL knockdown; CYP1B1 protein levels were assessed by western blotting at specified time points. H and I Treatment of 786O and ACHN cells with VHL overexpression or knockdown plasmids, followed by exposure to DMSO, chloroquine (CQ, 10 μM), or MG132 (20 μM) for 8 hours; CYP1B1 protein levels were analysed by western blotting. J and K Immunoprecipitation of cells with anti-Flag CYP1B1 antibodies, followed by western blotting with the indicated antibodies (ns, not significant; **, p < 0.01).
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    pVHL mediates the ubiquitination and degradation of CYP1B1. A and B Assessment of CYP1B1 mRNA and protein levels in ACHN cells with stable VHL knockdown and 786O and <t>OSRC2</t> cells with stable VHL overexpression via western blotting and RT‒qPCR. C Coimmunoprecipitation of ACHN cell whole-cell lysates with CYP1B1, pVHL or IgG antibodies, followed by western blotting with the respective antibodies. D 293T and 786O cells were transfected with Flag-CYP1B1 and His-pVHL plasmids. Coimmunoprecipitation with CYP1B1, pVHL or IgG antibodies was followed by western blotting with the indicated antibodies. E GST affinity-isolation assays with purified tagged pVHL and CYP1B1 proteins, followed by western blotting analysis. F Cotransfection of 293T and ACHN cells with VHL and CYP1B1 overexpression plasmids and subsequent immunofluorescence staining with anti-pVHL and anti-CYP1B1 antibodies. Scale bar: 10 μm. G Protein synthesis inhibition via CHX (10 μM) in 786O cells with VHL overexpression and ACHN cells with VHL knockdown; CYP1B1 protein levels were assessed by western blotting at specified time points. H and I Treatment of 786O and ACHN cells with VHL overexpression or knockdown plasmids, followed by exposure to DMSO, chloroquine (CQ, 10 μM), or MG132 (20 μM) for 8 hours; CYP1B1 protein levels were analysed by western blotting. J and K Immunoprecipitation of cells with anti-Flag CYP1B1 antibodies, followed by western blotting with the indicated antibodies (ns, not significant; **, p < 0.01).
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    control osrc2 cells  (Thermo Fisher)
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    Thermo Fisher control osrc2 cells
    ZNF582-AS1 expression was regulated by DNA methylation in ccRCC. a Detection of CpG islands in ZNF582-AS1 promoter and design of MSP primers. The horizontal axis of the curved lines represents the input sequence of ZNF582-AS1, and the vertical axis of the curved lines represents GC percentage. TSS: Transcription Start Sites. b MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC cell lines. c MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC tissues. d Detection of 38 CpG sites in ZNF582-AS1 promoter. e Quantitative detection of DNA methylation level of 38 CpG sites in ZNF582-AS1 promoter using Sequenom MassARRAY quantitative DNA methylation analysis. f and g Comparison of the DNA methylation levels of 38 CpG sites in ccRCC and adjacent normal renal tissues. h Treatment with 5-aza-dC and TSA demethylated ZNF582-AS1 promoter and increased ZNF582-AS1 expression in <t>OSRC2</t> and Caki-1 cells. T refers to Tumor tissue of ccRCC, N refers to Adjacent normal kidney tissue. M = Methylated, U = Unmethylated
    Control Osrc2 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pVHL mediates the ubiquitination and degradation of CYP1B1. A and B Assessment of CYP1B1 mRNA and protein levels in ACHN cells with stable VHL knockdown and 786O and OSRC2 cells with stable VHL overexpression via western blotting and RT‒qPCR. C Coimmunoprecipitation of ACHN cell whole-cell lysates with CYP1B1, pVHL or IgG antibodies, followed by western blotting with the respective antibodies. D 293T and 786O cells were transfected with Flag-CYP1B1 and His-pVHL plasmids. Coimmunoprecipitation with CYP1B1, pVHL or IgG antibodies was followed by western blotting with the indicated antibodies. E GST affinity-isolation assays with purified tagged pVHL and CYP1B1 proteins, followed by western blotting analysis. F Cotransfection of 293T and ACHN cells with VHL and CYP1B1 overexpression plasmids and subsequent immunofluorescence staining with anti-pVHL and anti-CYP1B1 antibodies. Scale bar: 10 μm. G Protein synthesis inhibition via CHX (10 μM) in 786O cells with VHL overexpression and ACHN cells with VHL knockdown; CYP1B1 protein levels were assessed by western blotting at specified time points. H and I Treatment of 786O and ACHN cells with VHL overexpression or knockdown plasmids, followed by exposure to DMSO, chloroquine (CQ, 10 μM), or MG132 (20 μM) for 8 hours; CYP1B1 protein levels were analysed by western blotting. J and K Immunoprecipitation of cells with anti-Flag CYP1B1 antibodies, followed by western blotting with the indicated antibodies (ns, not significant; **, p < 0.01).

    Journal: Neoplasia (New York, N.Y.)

    Article Title: CYP1B1 promotes angiogenesis and sunitinib resistance in clear cell renal cell carcinoma via USP5-mediated HIF2α deubiquitination

    doi: 10.1016/j.neo.2025.101186

    Figure Lengend Snippet: pVHL mediates the ubiquitination and degradation of CYP1B1. A and B Assessment of CYP1B1 mRNA and protein levels in ACHN cells with stable VHL knockdown and 786O and OSRC2 cells with stable VHL overexpression via western blotting and RT‒qPCR. C Coimmunoprecipitation of ACHN cell whole-cell lysates with CYP1B1, pVHL or IgG antibodies, followed by western blotting with the respective antibodies. D 293T and 786O cells were transfected with Flag-CYP1B1 and His-pVHL plasmids. Coimmunoprecipitation with CYP1B1, pVHL or IgG antibodies was followed by western blotting with the indicated antibodies. E GST affinity-isolation assays with purified tagged pVHL and CYP1B1 proteins, followed by western blotting analysis. F Cotransfection of 293T and ACHN cells with VHL and CYP1B1 overexpression plasmids and subsequent immunofluorescence staining with anti-pVHL and anti-CYP1B1 antibodies. Scale bar: 10 μm. G Protein synthesis inhibition via CHX (10 μM) in 786O cells with VHL overexpression and ACHN cells with VHL knockdown; CYP1B1 protein levels were assessed by western blotting at specified time points. H and I Treatment of 786O and ACHN cells with VHL overexpression or knockdown plasmids, followed by exposure to DMSO, chloroquine (CQ, 10 μM), or MG132 (20 μM) for 8 hours; CYP1B1 protein levels were analysed by western blotting. J and K Immunoprecipitation of cells with anti-Flag CYP1B1 antibodies, followed by western blotting with the indicated antibodies (ns, not significant; **, p < 0.01).

    Article Snippet: The OSRC2 cell line was acquired from ProCell (Wuhan, China) and maintained in RPMI-1640 medium enriched with 10 % FBS (Boster, Wuhan, China).

    Techniques: Ubiquitin Proteomics, Knockdown, Over Expression, Western Blot, Transfection, Isolation, Purification, Cotransfection, Immunofluorescence, Staining, Inhibition, Immunoprecipitation

    CYP1B1 regulates HIF2α protein stability through the ubiquitin–proteasome pathway. A Immunoprecipitation of 293T and 786O cell lysates with CYP1B1 or HIF2α antibodies, followed by western blotting with the indicated antibodies. B The purified tagged proteins were subjected to GST pull-down assays with the indicated antibodies. C Confocal microscopy analysis of HIF2α and CYP1B1 colocalization in 786O and OSRC2 cells. Scale bar: 50 μm. D Protein synthesis inhibition in 786O cells overexpressing CYP1B1 and SU-R 786O cells with CYP1B1 knockdown via CHX (10 μM); HIF2α protein levels were determined by western blotting at specified time points. E and F Treatment of 786O cells overexpressing CYP1B1 and SU-R 786O cells with CYP1B1 knockdown with DMSO, chloroquine (10 μM), or MG132 (20 μM) for 8 hours; HIF2α protein levels were analysed by western blotting. G Immunoprecipitation of the indicated cell lysates with anti-Flag antibodies, followed by western blotting with the indicated antibodies. H Venn diagram illustrating potential deubiquitinating enzymes that regulate the degradation of ubiquitinated HIF2α. I Co-IP assays in 786O cells were performed to assess the interaction between the five deubiquitinating enzymes and HIF2α. J The purified tagged proteins were subjected to GST pull-down assays with the indicated antibodies (ns, not significant; **, p < 0.01; ***, p < 0.001).

    Journal: Neoplasia (New York, N.Y.)

    Article Title: CYP1B1 promotes angiogenesis and sunitinib resistance in clear cell renal cell carcinoma via USP5-mediated HIF2α deubiquitination

    doi: 10.1016/j.neo.2025.101186

    Figure Lengend Snippet: CYP1B1 regulates HIF2α protein stability through the ubiquitin–proteasome pathway. A Immunoprecipitation of 293T and 786O cell lysates with CYP1B1 or HIF2α antibodies, followed by western blotting with the indicated antibodies. B The purified tagged proteins were subjected to GST pull-down assays with the indicated antibodies. C Confocal microscopy analysis of HIF2α and CYP1B1 colocalization in 786O and OSRC2 cells. Scale bar: 50 μm. D Protein synthesis inhibition in 786O cells overexpressing CYP1B1 and SU-R 786O cells with CYP1B1 knockdown via CHX (10 μM); HIF2α protein levels were determined by western blotting at specified time points. E and F Treatment of 786O cells overexpressing CYP1B1 and SU-R 786O cells with CYP1B1 knockdown with DMSO, chloroquine (10 μM), or MG132 (20 μM) for 8 hours; HIF2α protein levels were analysed by western blotting. G Immunoprecipitation of the indicated cell lysates with anti-Flag antibodies, followed by western blotting with the indicated antibodies. H Venn diagram illustrating potential deubiquitinating enzymes that regulate the degradation of ubiquitinated HIF2α. I Co-IP assays in 786O cells were performed to assess the interaction between the five deubiquitinating enzymes and HIF2α. J The purified tagged proteins were subjected to GST pull-down assays with the indicated antibodies (ns, not significant; **, p < 0.01; ***, p < 0.001).

    Article Snippet: The OSRC2 cell line was acquired from ProCell (Wuhan, China) and maintained in RPMI-1640 medium enriched with 10 % FBS (Boster, Wuhan, China).

    Techniques: Ubiquitin Proteomics, Immunoprecipitation, Western Blot, Purification, Confocal Microscopy, Inhibition, Knockdown, Co-Immunoprecipitation Assay

    ZNF582-AS1 expression was regulated by DNA methylation in ccRCC. a Detection of CpG islands in ZNF582-AS1 promoter and design of MSP primers. The horizontal axis of the curved lines represents the input sequence of ZNF582-AS1, and the vertical axis of the curved lines represents GC percentage. TSS: Transcription Start Sites. b MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC cell lines. c MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC tissues. d Detection of 38 CpG sites in ZNF582-AS1 promoter. e Quantitative detection of DNA methylation level of 38 CpG sites in ZNF582-AS1 promoter using Sequenom MassARRAY quantitative DNA methylation analysis. f and g Comparison of the DNA methylation levels of 38 CpG sites in ccRCC and adjacent normal renal tissues. h Treatment with 5-aza-dC and TSA demethylated ZNF582-AS1 promoter and increased ZNF582-AS1 expression in OSRC2 and Caki-1 cells. T refers to Tumor tissue of ccRCC, N refers to Adjacent normal kidney tissue. M = Methylated, U = Unmethylated

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

    doi: 10.1186/s13046-021-01889-8

    Figure Lengend Snippet: ZNF582-AS1 expression was regulated by DNA methylation in ccRCC. a Detection of CpG islands in ZNF582-AS1 promoter and design of MSP primers. The horizontal axis of the curved lines represents the input sequence of ZNF582-AS1, and the vertical axis of the curved lines represents GC percentage. TSS: Transcription Start Sites. b MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC cell lines. c MSP analysis of ZNF582-AS1 promoter DNA methylation status in ccRCC tissues. d Detection of 38 CpG sites in ZNF582-AS1 promoter. e Quantitative detection of DNA methylation level of 38 CpG sites in ZNF582-AS1 promoter using Sequenom MassARRAY quantitative DNA methylation analysis. f and g Comparison of the DNA methylation levels of 38 CpG sites in ccRCC and adjacent normal renal tissues. h Treatment with 5-aza-dC and TSA demethylated ZNF582-AS1 promoter and increased ZNF582-AS1 expression in OSRC2 and Caki-1 cells. T refers to Tumor tissue of ccRCC, N refers to Adjacent normal kidney tissue. M = Methylated, U = Unmethylated

    Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

    Techniques: Expressing, DNA Methylation Assay, Sequencing, Comparison, Methylation

    ZNF582-AS1 overexpression attenuated cell proliferation and induced cell apoptosis in vitro and in vivo. a and b EdU assay and c CCK-8 assay determined cell proliferation of ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells comparing with their control cells. d and e TUNEL assay and f and g flow cytometry determined cell apoptosis of ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells comparing with their control cells. h Tumors collected from mice are shown. i and j Tumor volume curves and tumor weights of the ZNF582-AS1-overexpressed and control groups were measured and compared. (k and l) The cell proliferation and apoptosis of tumors were examined

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

    doi: 10.1186/s13046-021-01889-8

    Figure Lengend Snippet: ZNF582-AS1 overexpression attenuated cell proliferation and induced cell apoptosis in vitro and in vivo. a and b EdU assay and c CCK-8 assay determined cell proliferation of ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells comparing with their control cells. d and e TUNEL assay and f and g flow cytometry determined cell apoptosis of ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells comparing with their control cells. h Tumors collected from mice are shown. i and j Tumor volume curves and tumor weights of the ZNF582-AS1-overexpressed and control groups were measured and compared. (k and l) The cell proliferation and apoptosis of tumors were examined

    Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

    Techniques: Over Expression, In Vitro, In Vivo, EdU Assay, CCK-8 Assay, Control, TUNEL Assay, Flow Cytometry

    ZNF582-AS1 overexpression inhibited cell migratory and invasive ability in vitro and in vivo. a and b Wound healing assay determined the migratory distances of ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells and their control cells. c and d Transwell migration and e and f invasion assays determined the migratory and invasive abilities of ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells and their control cells. g and h The luciferase signals in the ZNF582-AS1-overexpressed group were remarkably lower than those in the control group. i There was no significant difference between the mice weight of ZNF582-AS1-overexpressed and control group. j and k The number and size of pulmonary metastases in the ZNF582-AS1-overexpressed group were significantly reduced compared with those in the control group. l and m ZNF582-AS1 overexpression increased E-cadherin expression and decreased N-cadherin expression in pulmonary metastases

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

    doi: 10.1186/s13046-021-01889-8

    Figure Lengend Snippet: ZNF582-AS1 overexpression inhibited cell migratory and invasive ability in vitro and in vivo. a and b Wound healing assay determined the migratory distances of ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells and their control cells. c and d Transwell migration and e and f invasion assays determined the migratory and invasive abilities of ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells and their control cells. g and h The luciferase signals in the ZNF582-AS1-overexpressed group were remarkably lower than those in the control group. i There was no significant difference between the mice weight of ZNF582-AS1-overexpressed and control group. j and k The number and size of pulmonary metastases in the ZNF582-AS1-overexpressed group were significantly reduced compared with those in the control group. l and m ZNF582-AS1 overexpression increased E-cadherin expression and decreased N-cadherin expression in pulmonary metastases

    Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

    Techniques: Over Expression, In Vitro, In Vivo, Wound Healing Assay, Control, Migration, Luciferase, Expressing

    ZNF582-AS1 overexpression decreased rRNA adenine N(6)-methyltransferase A8K0B9 expression. a The overexpression efficiency of ZNF582-AS1 in OSRC2 cells. b iTRAQ results showed that 69 proteins were downregulated and 75 proteins were upregulated in ZNF582-AS1-overexpressed OSRC2 cells compared with control OSRC2 cells. c The expression of these 144 differently expressed proteins in ZNF582-AS1-overexpressed OSRC2 cells and control cells. d Biological Process GO term enrichment analysis of the complete 144 statistically significant proteins. e and f Validation of the presence of A8K0B9 protein. g A8K0B9 expression was decreased in ZNF582-AS1-overexpressed OSRC2 cells compared with control cells. h Changes of A8K0B9 protein level in OSRC2 cells after transient transfection of different doses of ZNF582-AS1 overexpression plasmid. i Estimation of the binding propensity of A8K0B9-ZNF582-AS1 pair. j RNA pull-down assay indicated that A8K0B9 protein bound specifically to ZNF582-AS1

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

    doi: 10.1186/s13046-021-01889-8

    Figure Lengend Snippet: ZNF582-AS1 overexpression decreased rRNA adenine N(6)-methyltransferase A8K0B9 expression. a The overexpression efficiency of ZNF582-AS1 in OSRC2 cells. b iTRAQ results showed that 69 proteins were downregulated and 75 proteins were upregulated in ZNF582-AS1-overexpressed OSRC2 cells compared with control OSRC2 cells. c The expression of these 144 differently expressed proteins in ZNF582-AS1-overexpressed OSRC2 cells and control cells. d Biological Process GO term enrichment analysis of the complete 144 statistically significant proteins. e and f Validation of the presence of A8K0B9 protein. g A8K0B9 expression was decreased in ZNF582-AS1-overexpressed OSRC2 cells compared with control cells. h Changes of A8K0B9 protein level in OSRC2 cells after transient transfection of different doses of ZNF582-AS1 overexpression plasmid. i Estimation of the binding propensity of A8K0B9-ZNF582-AS1 pair. j RNA pull-down assay indicated that A8K0B9 protein bound specifically to ZNF582-AS1

    Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

    Techniques: Over Expression, Expressing, Multiplex sample analysis, Control, Biomarker Discovery, Transfection, Plasmid Preparation, Binding Assay, Pull Down Assay

    ZNF582-AS1 regulated the N(6)-methyladenosine modification of MT-RNR1 by modulating A8K0B9. a Negative and positive controls of rRNA MeRIP-seq. b The analysis results of rRNA MeRIP-seq. c The expression of MT-RNR1, MT-RNR2 and RNA28SN5 in ZNF582-AS1-overexpressed and control OSRC2 cells. d and e MT-RNR1 expression was negatively associated with ZNF582-AS1 expression ( n = 530), and lower MT-RNR1 expression was related to longer OS (n = 530) based on TCGA-KIRC data. According to the median cutoff of MT-RNR1 expression, patients were divided into two groups for survival analysis. f The expression of MT-RNR1 in ccRCC cell lines. g Analysis of m6A motif DRACH (D = A, G or U; R = A or G; H = A, U or C) in MT-RNR1 sequence. h MeRIP-qPCR determined the methylation level of MT-RNR1 in ZNF582-AS1-overexpressed OSRC2 cells and control cells. i RIP-RT-qPCR showed that A8K0B9 protein had a certain binding ability with MT-RNR1 in OSRC2 cells

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

    doi: 10.1186/s13046-021-01889-8

    Figure Lengend Snippet: ZNF582-AS1 regulated the N(6)-methyladenosine modification of MT-RNR1 by modulating A8K0B9. a Negative and positive controls of rRNA MeRIP-seq. b The analysis results of rRNA MeRIP-seq. c The expression of MT-RNR1, MT-RNR2 and RNA28SN5 in ZNF582-AS1-overexpressed and control OSRC2 cells. d and e MT-RNR1 expression was negatively associated with ZNF582-AS1 expression ( n = 530), and lower MT-RNR1 expression was related to longer OS (n = 530) based on TCGA-KIRC data. According to the median cutoff of MT-RNR1 expression, patients were divided into two groups for survival analysis. f The expression of MT-RNR1 in ccRCC cell lines. g Analysis of m6A motif DRACH (D = A, G or U; R = A or G; H = A, U or C) in MT-RNR1 sequence. h MeRIP-qPCR determined the methylation level of MT-RNR1 in ZNF582-AS1-overexpressed OSRC2 cells and control cells. i RIP-RT-qPCR showed that A8K0B9 protein had a certain binding ability with MT-RNR1 in OSRC2 cells

    Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

    Techniques: Modification, Expressing, Control, Sequencing, Methylation, Quantitative RT-PCR, Binding Assay

    ZNF582-AS1 overexpression decreased MT-CO2 expression by regulating MT-RNR1. a and b MT-CO2 expression was significantly positively correlated with MT-RNR1 expression (n = 530), and lower MT-CO2 expression was associated with longer OS (n = 530) based on TCGA-KIRC data. According to the median cutoff of MT-CO2 expression, patients were divided into two groups for survival analysis. c Based on the iTRAQ results, the expression level of MT-CO2 (A0A0P0C1B5) protein was decreased in ZNF582-AS1-overexpressed OSRC2 cells compared with control OSRC2 cells. d and e MT-CO2, Bcl-2 and N-cadherin protein expression was decreased and Cleaved Caspase 3 and E-cadherin protein expression was increased in ZNF582-AS1-overexpressed OSRC2 cells compared with control cells. f and g The ROS level in ZNF582-AS1-overexpressed OSRC2 cells was increased. h The overexpression efficiency of MT-RNR1 in ZNF582-AS1-overexpressed OSRC2 cells. i and j MT-CO2, Bcl-2 and N-cadherin protein expression was increased and Cleaved Caspase 3 and E-cadherin protein expression was decreased in MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 cells compared with control cells. k and l The ROS level in MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 cells was decreased

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

    doi: 10.1186/s13046-021-01889-8

    Figure Lengend Snippet: ZNF582-AS1 overexpression decreased MT-CO2 expression by regulating MT-RNR1. a and b MT-CO2 expression was significantly positively correlated with MT-RNR1 expression (n = 530), and lower MT-CO2 expression was associated with longer OS (n = 530) based on TCGA-KIRC data. According to the median cutoff of MT-CO2 expression, patients were divided into two groups for survival analysis. c Based on the iTRAQ results, the expression level of MT-CO2 (A0A0P0C1B5) protein was decreased in ZNF582-AS1-overexpressed OSRC2 cells compared with control OSRC2 cells. d and e MT-CO2, Bcl-2 and N-cadherin protein expression was decreased and Cleaved Caspase 3 and E-cadherin protein expression was increased in ZNF582-AS1-overexpressed OSRC2 cells compared with control cells. f and g The ROS level in ZNF582-AS1-overexpressed OSRC2 cells was increased. h The overexpression efficiency of MT-RNR1 in ZNF582-AS1-overexpressed OSRC2 cells. i and j MT-CO2, Bcl-2 and N-cadherin protein expression was increased and Cleaved Caspase 3 and E-cadherin protein expression was decreased in MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 cells compared with control cells. k and l The ROS level in MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 cells was decreased

    Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

    Techniques: Over Expression, Expressing, Multiplex sample analysis, Control

    MT-RNR1 overexpression reversed inhibited cell proliferation and increased cell apoptosis caused by ZNF582-AS1 overexpression. a and b EdU assay and c CCK-8 assay determined cell proliferation of MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells comparing with their control cells. d and e TUNEL assay and f and g flow cytometry determined cell apoptosis of MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells comparing with their control cells. h Tumors collected from mice are shown. i and j Tumor volume curves and tumor weights of the MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control groups were measured and compared. k and l The cell proliferation and apoptosis of tumor were examined

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

    doi: 10.1186/s13046-021-01889-8

    Figure Lengend Snippet: MT-RNR1 overexpression reversed inhibited cell proliferation and increased cell apoptosis caused by ZNF582-AS1 overexpression. a and b EdU assay and c CCK-8 assay determined cell proliferation of MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells comparing with their control cells. d and e TUNEL assay and f and g flow cytometry determined cell apoptosis of MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells comparing with their control cells. h Tumors collected from mice are shown. i and j Tumor volume curves and tumor weights of the MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control groups were measured and compared. k and l The cell proliferation and apoptosis of tumor were examined

    Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

    Techniques: Over Expression, EdU Assay, CCK-8 Assay, Control, TUNEL Assay, Flow Cytometry

    MT-RNR1 overexpression reversed inhibited cell migratory and invasive ability caused by ZNF582-AS1 overexpression. a and b Wound healing assay determined the migratory distances of MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells and their control cells. c and d Transwell migration and e and f invasion assays determined the migratory and invasive abilities of MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells and their control cells. g and h The luciferase signals in the MT-RNR1-overexpressed-ZNF582-AS1-overexpressed group were remarkably higher than those in the ZNF582-AS1-overexpressed group. i There was no significant difference between the mouse weight of the two treatment group. j and k The number and size of pulmonary metastases in the MT-RNR1-overexpressed-ZNF582-AS1-overexpressed group were significantly increased compared with those in the ZNF582-AS1-overexpressed group. l and m MT-RNR1 overexpression decreased E-cadherin expression and increased N-cadherin expression in pulmonary metastases

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1

    doi: 10.1186/s13046-021-01889-8

    Figure Lengend Snippet: MT-RNR1 overexpression reversed inhibited cell migratory and invasive ability caused by ZNF582-AS1 overexpression. a and b Wound healing assay determined the migratory distances of MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells and their control cells. c and d Transwell migration and e and f invasion assays determined the migratory and invasive abilities of MT-RNR1-overexpressed-ZNF582-AS1-overexpressed OSRC2 and Caki-1 cells and their control cells. g and h The luciferase signals in the MT-RNR1-overexpressed-ZNF582-AS1-overexpressed group were remarkably higher than those in the ZNF582-AS1-overexpressed group. i There was no significant difference between the mouse weight of the two treatment group. j and k The number and size of pulmonary metastases in the MT-RNR1-overexpressed-ZNF582-AS1-overexpressed group were significantly increased compared with those in the ZNF582-AS1-overexpressed group. l and m MT-RNR1 overexpression decreased E-cadherin expression and increased N-cadherin expression in pulmonary metastases

    Article Snippet: Approximately 5 × 10 6 ZNF582-AS1-overexpressed, MT-RNR1-overexpressed-ZNF582-AS1-overexpressed and control OSRC2 cells suspended in 100 μL Hank’s Balanced Salt Solution (Thermo Fisher Scientific, USA) were mixed with Matrigel (1:1, Corning Inc., USA).

    Techniques: Over Expression, Wound Healing Assay, Control, Migration, Luciferase, Expressing