renal cancer cells  (ATCC)


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    ATCC renal cancer cells
    Renal Cancer Cells, supplied by ATCC, 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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    htb 44 rrid cvcl 1056  (ATCC)


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    ATCC htb 44 rrid cvcl 1056
    Htb 44 Rrid Cvcl 1056, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a498 htb 44  (ATCC)


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    ATCC a498 htb 44
    A498 Htb 44, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a498 htb 44  (ATCC)


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    ATCC a498 htb 44
    A498 Htb 44, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a498 htb 44  (ATCC)


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    ATCC a498 htb 44
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    a498 htb 44 cell lines  (ATCC)


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    ATCC a498 htb 44 cell lines
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    renal cancer cells  (ATCC)


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    ATCC renal cancer cells
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    786 o  (ATCC)


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    ATCC 786 o
    Effects of luteolin on cell viability. <t>786-O</t> cells were treated with various concentrations of luteolin. Representative images of phase contrast were obtained 24 h after treatment. Scale bar = 60 μ m (a). 786-O cells were treated with various concentrations of luteolin over time. Cell viability was determined by MTS reduction assay (b). 786-O cells were treated with various concentrations of luteolin for 24 h. The cells were harvested and processed by flow-cytometric analysis. The percentage of subG1 population is shown (c). Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (d). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (e). ** P < 0.01 versus medium control, n = 4.
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    1) Product Images from "Induction of Apoptosis by Luteolin Involving Akt Inactivation in Human 786-O Renal Cell Carcinoma Cells"

    Article Title: Induction of Apoptosis by Luteolin Involving Akt Inactivation in Human 786-O Renal Cell Carcinoma Cells

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    doi: 10.1155/2013/109105

    Effects of luteolin on cell viability. 786-O cells were treated with various concentrations of luteolin. Representative images of phase contrast were obtained 24 h after treatment. Scale bar = 60 μ m (a). 786-O cells were treated with various concentrations of luteolin over time. Cell viability was determined by MTS reduction assay (b). 786-O cells were treated with various concentrations of luteolin for 24 h. The cells were harvested and processed by flow-cytometric analysis. The percentage of subG1 population is shown (c). Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (d). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (e). ** P < 0.01 versus medium control, n = 4.
    Figure Legend Snippet: Effects of luteolin on cell viability. 786-O cells were treated with various concentrations of luteolin. Representative images of phase contrast were obtained 24 h after treatment. Scale bar = 60 μ m (a). 786-O cells were treated with various concentrations of luteolin over time. Cell viability was determined by MTS reduction assay (b). 786-O cells were treated with various concentrations of luteolin for 24 h. The cells were harvested and processed by flow-cytometric analysis. The percentage of subG1 population is shown (c). Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (d). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (e). ** P < 0.01 versus medium control, n = 4.

    Techniques Used: Isolation, Western Blot, Caspase-3 Assay

    Role of Akt. 786-O cells were treated with various concentrations of LY294002 for 24 h. Cell viability was determined by MTS reduction assay (a). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit. ** P < 0.01 versus medium control, n = 4 (b). Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (c). 786-O cells were treated with various concentrations of luteolin for 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (d). Proteins obtained from medium- and luteolin (40 μ M)-treated cells were immunoprecipitated by anti-Ask1 antibody, and the immunoprecipitates were subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (e).
    Figure Legend Snippet: Role of Akt. 786-O cells were treated with various concentrations of LY294002 for 24 h. Cell viability was determined by MTS reduction assay (a). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit. ** P < 0.01 versus medium control, n = 4 (b). Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (c). 786-O cells were treated with various concentrations of luteolin for 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (d). Proteins obtained from medium- and luteolin (40 μ M)-treated cells were immunoprecipitated by anti-Ask1 antibody, and the immunoprecipitates were subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (e).

    Techniques Used: Isolation, Caspase-3 Assay, Western Blot, Immunoprecipitation

    Effects of luteolin on MAPKs and Akt. 786-O cells were treated with various concentrations of luteolin for 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (a). 786-O cells were pretreated with medium, SB203580, SP600125, U0126, or insulin for 1 h and then were treated with luteolin for additional 24 h. Cell viability was determined by MTS reduction assay (b). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (c). ** P < 0.01 versus medium control and ## P < 0.01 versus luteolin control, n = 4.
    Figure Legend Snippet: Effects of luteolin on MAPKs and Akt. 786-O cells were treated with various concentrations of luteolin for 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (a). 786-O cells were pretreated with medium, SB203580, SP600125, U0126, or insulin for 1 h and then were treated with luteolin for additional 24 h. Cell viability was determined by MTS reduction assay (b). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (c). ** P < 0.01 versus medium control and ## P < 0.01 versus luteolin control, n = 4.

    Techniques Used: Isolation, Western Blot, Caspase-3 Assay

    Role of HSP90. 786-O cells were treated with various concentrations of luteolin for 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown. An additional band of HSP90 was indicated by arrow (a). 786-O cells were treated with various concentrations of 17-AAG for 24 h. Cell viability was determined by MTS reduction assay (b). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit. ** P < 0.01 versus medium control, n = 4 (c). Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (d). Proteins obtained from medium- and luteolin (40 μ M)-treated cells were immunoprecipitated by anti-HSP90 antibody, and the immunoprecipitates were subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (e).
    Figure Legend Snippet: Role of HSP90. 786-O cells were treated with various concentrations of luteolin for 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown. An additional band of HSP90 was indicated by arrow (a). 786-O cells were treated with various concentrations of 17-AAG for 24 h. Cell viability was determined by MTS reduction assay (b). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit. ** P < 0.01 versus medium control, n = 4 (c). Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (d). Proteins obtained from medium- and luteolin (40 μ M)-treated cells were immunoprecipitated by anti-HSP90 antibody, and the immunoprecipitates were subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (e).

    Techniques Used: Isolation, Western Blot, Caspase-3 Assay, Immunoprecipitation

    Effects of agents on HSP90 cleavage. 786-O cells were treated with luteolin (0 and 40 μ M) for 1 h, 3 h, and 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (a). 786-O cells were treated with various concentrations of luteolin for 1 h and 24 h. Protein extracts were isolated and subjected to enzymatic assay for measurement of PP2A. ** P < 0.01 versus medium control, n = 4 (b). 786-O cells were pretreated with medium, SB203580, SP600125, LY294002, or Z-VAD-FMK for 1 h and then were treated with luteolin for additional 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. An additional band of HSP90 was indicated by arrow. One of four independent experiments is shown (c).
    Figure Legend Snippet: Effects of agents on HSP90 cleavage. 786-O cells were treated with luteolin (0 and 40 μ M) for 1 h, 3 h, and 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (a). 786-O cells were treated with various concentrations of luteolin for 1 h and 24 h. Protein extracts were isolated and subjected to enzymatic assay for measurement of PP2A. ** P < 0.01 versus medium control, n = 4 (b). 786-O cells were pretreated with medium, SB203580, SP600125, LY294002, or Z-VAD-FMK for 1 h and then were treated with luteolin for additional 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. An additional band of HSP90 was indicated by arrow. One of four independent experiments is shown (c).

    Techniques Used: Isolation, Western Blot, Enzymatic Assay

    Effects of agents on cell viability. 786-O cells were treated with medium, luteolin, 17-AAG, or LY294002 alone, or in combinations for 24 h. Cell viability was determined by MTS reduction assay (a). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (b). ** P < 0.01 versus medium control, ## P < 0.01 versus luteolin control, && P < 0.01 versus luteolin/17-AAG, and %% P < 0.01 versus luteolin/LY294002, n = 4.
    Figure Legend Snippet: Effects of agents on cell viability. 786-O cells were treated with medium, luteolin, 17-AAG, or LY294002 alone, or in combinations for 24 h. Cell viability was determined by MTS reduction assay (a). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (b). ** P < 0.01 versus medium control, ## P < 0.01 versus luteolin control, && P < 0.01 versus luteolin/17-AAG, and %% P < 0.01 versus luteolin/LY294002, n = 4.

    Techniques Used: Isolation, Caspase-3 Assay

    possible signaling cascade of apoptosis elicited by luteolin in 786-O cells is proposed. This schematic diagram indicates the signaling molecules employed in mediating 786-O cell apoptosis after luteolin treatment. Some additional signaling molecules and cascades have been omitted for the sake of clarity.
    Figure Legend Snippet: possible signaling cascade of apoptosis elicited by luteolin in 786-O cells is proposed. This schematic diagram indicates the signaling molecules employed in mediating 786-O cell apoptosis after luteolin treatment. Some additional signaling molecules and cascades have been omitted for the sake of clarity.

    Techniques Used:

    a498  (ATCC)


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    ATCC a498
    Effects of luteolin on <t>A498</t> and ACHN cells. A498 and ACHN cells were treated with various concentrations of luteolin for 24 h. Cell viability was determined by MTS reduction assay (a). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (b). * P < 0.05 and ** P < 0.01 versus medium control, n = 4. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of three independent experiments is shown (c). An additional band of HSP90 was indicated by arrow.
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    1) Product Images from "Induction of Apoptosis by Luteolin Involving Akt Inactivation in Human 786-O Renal Cell Carcinoma Cells"

    Article Title: Induction of Apoptosis by Luteolin Involving Akt Inactivation in Human 786-O Renal Cell Carcinoma Cells

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    doi: 10.1155/2013/109105

    Effects of luteolin on A498 and ACHN cells. A498 and ACHN cells were treated with various concentrations of luteolin for 24 h. Cell viability was determined by MTS reduction assay (a). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (b). * P < 0.05 and ** P < 0.01 versus medium control, n = 4. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of three independent experiments is shown (c). An additional band of HSP90 was indicated by arrow.
    Figure Legend Snippet: Effects of luteolin on A498 and ACHN cells. A498 and ACHN cells were treated with various concentrations of luteolin for 24 h. Cell viability was determined by MTS reduction assay (a). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (b). * P < 0.05 and ** P < 0.01 versus medium control, n = 4. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of three independent experiments is shown (c). An additional band of HSP90 was indicated by arrow.

    Techniques Used: Isolation, Caspase-3 Assay, Western Blot

    a498  (ATCC)


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    ATCC a498
    Everolimus (RAD001) induces autophagy in RCCs. Notes: ( A , B ) 786-O and <t>A498</t> cell lines were treated by RAD001 with different concentrations (0, 0.01, 0.1, 1, 10, and 20 μmol/L) for 24 hours. ( A ) Immunoblotting showed that RAD001 induced autophagy indicators including upregulation of LC3-II/I and downregulation of p62 in a dose-dependent manner. ( B ) Quantification of the immunoblots. * P <0.05 versus 0 μmol/L group. n=3. ( C , D ) 786-O and A498 cell lines were treated by RAD001 for 0, 1, 2, 4, and 8 hours. ( C ) Immunoblotting showed that RAD001 induced autophagy in 786-O and A498 cells as time prolonged, which peaked at 8 hours. ( D ) Quantification of the immunoblots. * P <0.05 versus 0 hour group. n=3. Abbreviation: RCCs, renal cell carcinoma cells.
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    1) Product Images from "Attenuation of everolimus-induced cytotoxicity by a protective autophagic pathway involving ERK activation in renal cell carcinoma cells"

    Article Title: Attenuation of everolimus-induced cytotoxicity by a protective autophagic pathway involving ERK activation in renal cell carcinoma cells

    Journal: Drug Design, Development and Therapy

    doi: 10.2147/DDDT.S160557

    Everolimus (RAD001) induces autophagy in RCCs. Notes: ( A , B ) 786-O and A498 cell lines were treated by RAD001 with different concentrations (0, 0.01, 0.1, 1, 10, and 20 μmol/L) for 24 hours. ( A ) Immunoblotting showed that RAD001 induced autophagy indicators including upregulation of LC3-II/I and downregulation of p62 in a dose-dependent manner. ( B ) Quantification of the immunoblots. * P <0.05 versus 0 μmol/L group. n=3. ( C , D ) 786-O and A498 cell lines were treated by RAD001 for 0, 1, 2, 4, and 8 hours. ( C ) Immunoblotting showed that RAD001 induced autophagy in 786-O and A498 cells as time prolonged, which peaked at 8 hours. ( D ) Quantification of the immunoblots. * P <0.05 versus 0 hour group. n=3. Abbreviation: RCCs, renal cell carcinoma cells.
    Figure Legend Snippet: Everolimus (RAD001) induces autophagy in RCCs. Notes: ( A , B ) 786-O and A498 cell lines were treated by RAD001 with different concentrations (0, 0.01, 0.1, 1, 10, and 20 μmol/L) for 24 hours. ( A ) Immunoblotting showed that RAD001 induced autophagy indicators including upregulation of LC3-II/I and downregulation of p62 in a dose-dependent manner. ( B ) Quantification of the immunoblots. * P <0.05 versus 0 μmol/L group. n=3. ( C , D ) 786-O and A498 cell lines were treated by RAD001 for 0, 1, 2, 4, and 8 hours. ( C ) Immunoblotting showed that RAD001 induced autophagy in 786-O and A498 cells as time prolonged, which peaked at 8 hours. ( D ) Quantification of the immunoblots. * P <0.05 versus 0 hour group. n=3. Abbreviation: RCCs, renal cell carcinoma cells.

    Techniques Used: Western Blot

    Inhibition of autophagy enhances RAD001-induced cytotoxicity, RAD001-induced activation of ERK signaling pathway in RCCs. Notes: 786-O and A498 cells were pretreated with CQ (autophagy inhibitor) (10 mmol/L) for 30 minutes and then treated with RAD001 for 8 hours. ( A ) MTT assay showed that RAD001 impaired cell viability of 786-O and A498 cells in a dose-dependent manner. n=3. ( B ) Immunoblotting of LC3-II/I and p62 showed that CQ could restore the downregulation of p62, but would not impair the upregulation of LC3-II/I because CQ blocks autophagy at the lysosomal degradation step. ( C ) Densitometric analysis was performed to quantify the immunoblots. * P <0.05 versus control. # P <0.05 versus RAD001 group. n=3. ( D ) MTT assay displayed that autophagy inhibition by CQ promoted RAD001-induced cytotoxicity in 786-O and A498 cells. * P <0.05 versus control. # P <0.05 versus RAD001 group. n=3. ( E ) The apoptosis indicator cleave-PARP detected by immunoblot indicated that cells apoptosis was increased in 786-O and A498 cells after CQ intervention, compared to the RAD001 group. ( F ) Densitometric analysis was performed to quantify the immunoblots. * P <0.05 versus control group. # P <0.05 versus RAD001 group. n=3. 786-O and A498 cells were treated by RAD001 for 0, 1, 2, 4, and 8 hours. ( G ) Immunoblotting analyzed the JNK, p38, and ERK signaling molecules in 786-O and A498 cells after RAD001 treatment. ( H ) Quantification of the immunoblots. * P <0.05 versus 0 hour group. n=3. Abbreviations: CQ, chloroquine; Con, control; RCCs, renal cell carcinoma cells; ERK, extracellular signal-regulated kinase; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PARP, poly ADP ribose polymerase; JNK, c-Jun N-terminal kinase.
    Figure Legend Snippet: Inhibition of autophagy enhances RAD001-induced cytotoxicity, RAD001-induced activation of ERK signaling pathway in RCCs. Notes: 786-O and A498 cells were pretreated with CQ (autophagy inhibitor) (10 mmol/L) for 30 minutes and then treated with RAD001 for 8 hours. ( A ) MTT assay showed that RAD001 impaired cell viability of 786-O and A498 cells in a dose-dependent manner. n=3. ( B ) Immunoblotting of LC3-II/I and p62 showed that CQ could restore the downregulation of p62, but would not impair the upregulation of LC3-II/I because CQ blocks autophagy at the lysosomal degradation step. ( C ) Densitometric analysis was performed to quantify the immunoblots. * P <0.05 versus control. # P <0.05 versus RAD001 group. n=3. ( D ) MTT assay displayed that autophagy inhibition by CQ promoted RAD001-induced cytotoxicity in 786-O and A498 cells. * P <0.05 versus control. # P <0.05 versus RAD001 group. n=3. ( E ) The apoptosis indicator cleave-PARP detected by immunoblot indicated that cells apoptosis was increased in 786-O and A498 cells after CQ intervention, compared to the RAD001 group. ( F ) Densitometric analysis was performed to quantify the immunoblots. * P <0.05 versus control group. # P <0.05 versus RAD001 group. n=3. 786-O and A498 cells were treated by RAD001 for 0, 1, 2, 4, and 8 hours. ( G ) Immunoblotting analyzed the JNK, p38, and ERK signaling molecules in 786-O and A498 cells after RAD001 treatment. ( H ) Quantification of the immunoblots. * P <0.05 versus 0 hour group. n=3. Abbreviations: CQ, chloroquine; Con, control; RCCs, renal cell carcinoma cells; ERK, extracellular signal-regulated kinase; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PARP, poly ADP ribose polymerase; JNK, c-Jun N-terminal kinase.

    Techniques Used: Inhibition, Activation Assay, MTT Assay, Western Blot

    Selumetinib (AZD6244) significantly enhances RAD001-induced apoptosis of RCCs. Notes: 786-O and A498 cells were pretreated with AZD6244 (1 μmol/L) for 30 minutes and then treated with RAD001 for 8 hours. ( A ) Immunoblotting showed that AZD6244 significantly inhibited the activation of ERK pathway in 786-O and A498 cells. ( B ) Quantification of the immunoblots. ( C ) MTT assay detected the effect of AZD6244 on cell viability of 786-O and A498 cells, which suggested that AZD6244 effectively blunts RAD001-induced cell viability. ( D ) Immunoblotting demonstrates that AZD6244 could effectively restore upregulation of LC3-II/I and downregulation of p62 induced by RAD001 in 786-O and A498 cells. ( E ) Densitometric analysis was performed to quantify the immunoblots. ( F ) Immunoblot analysis indicated that combination treatment of RAD001 with AZD6244 could increase cleave-PARP level in 786-O and A498 cells, compared to the RAD001 group. ( G ) Densitometric analysis was performed to quantify the cleave-PARP immunoblots. ( H ) The difference of apoptosis determined by flow cytometry showed that AZD6244 significantly increased RAD001-induced cells apoptosis in 786-O and A498 cell lines. ( I ) Quantification of the apoptosis proportion. * P <0.05 versus control group. # P <0.05 versus RAD001 group. n=3. Abbreviations: RCCs, renal cell carcinoma cells; ERK, extracellular signal-regulated kinase; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PARP, poly ADP ribose polymerase; FITC, fluorescein isothiocyanate; PI, propidium iodide; RAD001, everolimus; AZD6244, selumetinib; Con, control.
    Figure Legend Snippet: Selumetinib (AZD6244) significantly enhances RAD001-induced apoptosis of RCCs. Notes: 786-O and A498 cells were pretreated with AZD6244 (1 μmol/L) for 30 minutes and then treated with RAD001 for 8 hours. ( A ) Immunoblotting showed that AZD6244 significantly inhibited the activation of ERK pathway in 786-O and A498 cells. ( B ) Quantification of the immunoblots. ( C ) MTT assay detected the effect of AZD6244 on cell viability of 786-O and A498 cells, which suggested that AZD6244 effectively blunts RAD001-induced cell viability. ( D ) Immunoblotting demonstrates that AZD6244 could effectively restore upregulation of LC3-II/I and downregulation of p62 induced by RAD001 in 786-O and A498 cells. ( E ) Densitometric analysis was performed to quantify the immunoblots. ( F ) Immunoblot analysis indicated that combination treatment of RAD001 with AZD6244 could increase cleave-PARP level in 786-O and A498 cells, compared to the RAD001 group. ( G ) Densitometric analysis was performed to quantify the cleave-PARP immunoblots. ( H ) The difference of apoptosis determined by flow cytometry showed that AZD6244 significantly increased RAD001-induced cells apoptosis in 786-O and A498 cell lines. ( I ) Quantification of the apoptosis proportion. * P <0.05 versus control group. # P <0.05 versus RAD001 group. n=3. Abbreviations: RCCs, renal cell carcinoma cells; ERK, extracellular signal-regulated kinase; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PARP, poly ADP ribose polymerase; FITC, fluorescein isothiocyanate; PI, propidium iodide; RAD001, everolimus; AZD6244, selumetinib; Con, control.

    Techniques Used: Western Blot, Activation Assay, MTT Assay, Flow Cytometry

    ERK mediated autophagy through upregulating Beclin-1 and p-Bcl-2 expression. Notes: ( A , B ) 786-O and A498 cells were treated by RAD001 for 0, 1, 2, 4, and 8 hours. ( A ) Immunoblotting showed that RAD001 induced upregulation of downstream signaling molecules of the ERK pathway including Beclin-1 and Bcl-2 in 786-O and A498 cells in a time-dependent manner. ( B ) Densitometric analysis was performed to quantify the immunoblots. * P <0.05 versus 0 hour group. n=3. ( C , D ) 786-O and A498 cells were pretreated with AZD6244 (1 μmol/L) for 30 minutes and then treated with RAD001 for 8 hours. ( C ) Immunoblotting showed that AZD6244 significantly decreased protein levels of Beclin-1 and Bcl-2 in 786-O and A498 cells. ( D ) Densitometric analysis was performed to quantify the immunoblots. * P <0.05 versus control group. # P <0.05 versus RAD001 group. n=3. Abbreviations: ERK, extracellular signal-regulated kinase; RAD001, everolimus.
    Figure Legend Snippet: ERK mediated autophagy through upregulating Beclin-1 and p-Bcl-2 expression. Notes: ( A , B ) 786-O and A498 cells were treated by RAD001 for 0, 1, 2, 4, and 8 hours. ( A ) Immunoblotting showed that RAD001 induced upregulation of downstream signaling molecules of the ERK pathway including Beclin-1 and Bcl-2 in 786-O and A498 cells in a time-dependent manner. ( B ) Densitometric analysis was performed to quantify the immunoblots. * P <0.05 versus 0 hour group. n=3. ( C , D ) 786-O and A498 cells were pretreated with AZD6244 (1 μmol/L) for 30 minutes and then treated with RAD001 for 8 hours. ( C ) Immunoblotting showed that AZD6244 significantly decreased protein levels of Beclin-1 and Bcl-2 in 786-O and A498 cells. ( D ) Densitometric analysis was performed to quantify the immunoblots. * P <0.05 versus control group. # P <0.05 versus RAD001 group. n=3. Abbreviations: ERK, extracellular signal-regulated kinase; RAD001, everolimus.

    Techniques Used: Expressing, Western Blot

    a498  (ATCC)


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    Effects of luteolin on cell viability. <t>786-O</t> cells were treated with various concentrations of luteolin. Representative images of phase contrast were obtained 24 h after treatment. Scale bar = 60 μ m (a). 786-O cells were treated with various concentrations of luteolin over time. Cell viability was determined by MTS reduction assay (b). 786-O cells were treated with various concentrations of luteolin for 24 h. The cells were harvested and processed by flow-cytometric analysis. The percentage of subG1 population is shown (c). Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (d). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (e). ** P < 0.01 versus medium control, n = 4.
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    Effects of luteolin on <t>A498</t> and ACHN cells. A498 and ACHN cells were treated with various concentrations of luteolin for 24 h. Cell viability was determined by MTS reduction assay (a). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (b). * P < 0.05 and ** P < 0.01 versus medium control, n = 4. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of three independent experiments is shown (c). An additional band of HSP90 was indicated by arrow.
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    Effects of luteolin on cell viability. 786-O cells were treated with various concentrations of luteolin. Representative images of phase contrast were obtained 24 h after treatment. Scale bar = 60 μ m (a). 786-O cells were treated with various concentrations of luteolin over time. Cell viability was determined by MTS reduction assay (b). 786-O cells were treated with various concentrations of luteolin for 24 h. The cells were harvested and processed by flow-cytometric analysis. The percentage of subG1 population is shown (c). Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (d). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (e). ** P < 0.01 versus medium control, n = 4.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Induction of Apoptosis by Luteolin Involving Akt Inactivation in Human 786-O Renal Cell Carcinoma Cells

    doi: 10.1155/2013/109105

    Figure Lengend Snippet: Effects of luteolin on cell viability. 786-O cells were treated with various concentrations of luteolin. Representative images of phase contrast were obtained 24 h after treatment. Scale bar = 60 μ m (a). 786-O cells were treated with various concentrations of luteolin over time. Cell viability was determined by MTS reduction assay (b). 786-O cells were treated with various concentrations of luteolin for 24 h. The cells were harvested and processed by flow-cytometric analysis. The percentage of subG1 population is shown (c). Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (d). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (e). ** P < 0.01 versus medium control, n = 4.

    Article Snippet: Human RCC cell lines, 786-O (ATCC CRL1932), A498 (ATCC HTB-44), and ACHN (ATCC CRL-1611) were cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μ g/mL streptomycin and were maintained in a humidified incubator with 5% CO 2 .

    Techniques: Isolation, Western Blot, Caspase-3 Assay

    Role of Akt. 786-O cells were treated with various concentrations of LY294002 for 24 h. Cell viability was determined by MTS reduction assay (a). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit. ** P < 0.01 versus medium control, n = 4 (b). Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (c). 786-O cells were treated with various concentrations of luteolin for 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (d). Proteins obtained from medium- and luteolin (40 μ M)-treated cells were immunoprecipitated by anti-Ask1 antibody, and the immunoprecipitates were subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (e).

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Induction of Apoptosis by Luteolin Involving Akt Inactivation in Human 786-O Renal Cell Carcinoma Cells

    doi: 10.1155/2013/109105

    Figure Lengend Snippet: Role of Akt. 786-O cells were treated with various concentrations of LY294002 for 24 h. Cell viability was determined by MTS reduction assay (a). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit. ** P < 0.01 versus medium control, n = 4 (b). Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (c). 786-O cells were treated with various concentrations of luteolin for 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (d). Proteins obtained from medium- and luteolin (40 μ M)-treated cells were immunoprecipitated by anti-Ask1 antibody, and the immunoprecipitates were subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (e).

    Article Snippet: Human RCC cell lines, 786-O (ATCC CRL1932), A498 (ATCC HTB-44), and ACHN (ATCC CRL-1611) were cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μ g/mL streptomycin and were maintained in a humidified incubator with 5% CO 2 .

    Techniques: Isolation, Caspase-3 Assay, Western Blot, Immunoprecipitation

    Effects of luteolin on MAPKs and Akt. 786-O cells were treated with various concentrations of luteolin for 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (a). 786-O cells were pretreated with medium, SB203580, SP600125, U0126, or insulin for 1 h and then were treated with luteolin for additional 24 h. Cell viability was determined by MTS reduction assay (b). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (c). ** P < 0.01 versus medium control and ## P < 0.01 versus luteolin control, n = 4.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Induction of Apoptosis by Luteolin Involving Akt Inactivation in Human 786-O Renal Cell Carcinoma Cells

    doi: 10.1155/2013/109105

    Figure Lengend Snippet: Effects of luteolin on MAPKs and Akt. 786-O cells were treated with various concentrations of luteolin for 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (a). 786-O cells were pretreated with medium, SB203580, SP600125, U0126, or insulin for 1 h and then were treated with luteolin for additional 24 h. Cell viability was determined by MTS reduction assay (b). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (c). ** P < 0.01 versus medium control and ## P < 0.01 versus luteolin control, n = 4.

    Article Snippet: Human RCC cell lines, 786-O (ATCC CRL1932), A498 (ATCC HTB-44), and ACHN (ATCC CRL-1611) were cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μ g/mL streptomycin and were maintained in a humidified incubator with 5% CO 2 .

    Techniques: Isolation, Western Blot, Caspase-3 Assay

    Role of HSP90. 786-O cells were treated with various concentrations of luteolin for 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown. An additional band of HSP90 was indicated by arrow (a). 786-O cells were treated with various concentrations of 17-AAG for 24 h. Cell viability was determined by MTS reduction assay (b). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit. ** P < 0.01 versus medium control, n = 4 (c). Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (d). Proteins obtained from medium- and luteolin (40 μ M)-treated cells were immunoprecipitated by anti-HSP90 antibody, and the immunoprecipitates were subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (e).

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Induction of Apoptosis by Luteolin Involving Akt Inactivation in Human 786-O Renal Cell Carcinoma Cells

    doi: 10.1155/2013/109105

    Figure Lengend Snippet: Role of HSP90. 786-O cells were treated with various concentrations of luteolin for 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown. An additional band of HSP90 was indicated by arrow (a). 786-O cells were treated with various concentrations of 17-AAG for 24 h. Cell viability was determined by MTS reduction assay (b). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit. ** P < 0.01 versus medium control, n = 4 (c). Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (d). Proteins obtained from medium- and luteolin (40 μ M)-treated cells were immunoprecipitated by anti-HSP90 antibody, and the immunoprecipitates were subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (e).

    Article Snippet: Human RCC cell lines, 786-O (ATCC CRL1932), A498 (ATCC HTB-44), and ACHN (ATCC CRL-1611) were cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μ g/mL streptomycin and were maintained in a humidified incubator with 5% CO 2 .

    Techniques: Isolation, Western Blot, Caspase-3 Assay, Immunoprecipitation

    Effects of agents on HSP90 cleavage. 786-O cells were treated with luteolin (0 and 40 μ M) for 1 h, 3 h, and 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (a). 786-O cells were treated with various concentrations of luteolin for 1 h and 24 h. Protein extracts were isolated and subjected to enzymatic assay for measurement of PP2A. ** P < 0.01 versus medium control, n = 4 (b). 786-O cells were pretreated with medium, SB203580, SP600125, LY294002, or Z-VAD-FMK for 1 h and then were treated with luteolin for additional 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. An additional band of HSP90 was indicated by arrow. One of four independent experiments is shown (c).

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Induction of Apoptosis by Luteolin Involving Akt Inactivation in Human 786-O Renal Cell Carcinoma Cells

    doi: 10.1155/2013/109105

    Figure Lengend Snippet: Effects of agents on HSP90 cleavage. 786-O cells were treated with luteolin (0 and 40 μ M) for 1 h, 3 h, and 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of four independent experiments is shown (a). 786-O cells were treated with various concentrations of luteolin for 1 h and 24 h. Protein extracts were isolated and subjected to enzymatic assay for measurement of PP2A. ** P < 0.01 versus medium control, n = 4 (b). 786-O cells were pretreated with medium, SB203580, SP600125, LY294002, or Z-VAD-FMK for 1 h and then were treated with luteolin for additional 24 h. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. An additional band of HSP90 was indicated by arrow. One of four independent experiments is shown (c).

    Article Snippet: Human RCC cell lines, 786-O (ATCC CRL1932), A498 (ATCC HTB-44), and ACHN (ATCC CRL-1611) were cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μ g/mL streptomycin and were maintained in a humidified incubator with 5% CO 2 .

    Techniques: Isolation, Western Blot, Enzymatic Assay

    Effects of agents on cell viability. 786-O cells were treated with medium, luteolin, 17-AAG, or LY294002 alone, or in combinations for 24 h. Cell viability was determined by MTS reduction assay (a). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (b). ** P < 0.01 versus medium control, ## P < 0.01 versus luteolin control, && P < 0.01 versus luteolin/17-AAG, and %% P < 0.01 versus luteolin/LY294002, n = 4.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Induction of Apoptosis by Luteolin Involving Akt Inactivation in Human 786-O Renal Cell Carcinoma Cells

    doi: 10.1155/2013/109105

    Figure Lengend Snippet: Effects of agents on cell viability. 786-O cells were treated with medium, luteolin, 17-AAG, or LY294002 alone, or in combinations for 24 h. Cell viability was determined by MTS reduction assay (a). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (b). ** P < 0.01 versus medium control, ## P < 0.01 versus luteolin control, && P < 0.01 versus luteolin/17-AAG, and %% P < 0.01 versus luteolin/LY294002, n = 4.

    Article Snippet: Human RCC cell lines, 786-O (ATCC CRL1932), A498 (ATCC HTB-44), and ACHN (ATCC CRL-1611) were cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μ g/mL streptomycin and were maintained in a humidified incubator with 5% CO 2 .

    Techniques: Isolation, Caspase-3 Assay

    possible signaling cascade of apoptosis elicited by luteolin in 786-O cells is proposed. This schematic diagram indicates the signaling molecules employed in mediating 786-O cell apoptosis after luteolin treatment. Some additional signaling molecules and cascades have been omitted for the sake of clarity.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Induction of Apoptosis by Luteolin Involving Akt Inactivation in Human 786-O Renal Cell Carcinoma Cells

    doi: 10.1155/2013/109105

    Figure Lengend Snippet: possible signaling cascade of apoptosis elicited by luteolin in 786-O cells is proposed. This schematic diagram indicates the signaling molecules employed in mediating 786-O cell apoptosis after luteolin treatment. Some additional signaling molecules and cascades have been omitted for the sake of clarity.

    Article Snippet: Human RCC cell lines, 786-O (ATCC CRL1932), A498 (ATCC HTB-44), and ACHN (ATCC CRL-1611) were cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μ g/mL streptomycin and were maintained in a humidified incubator with 5% CO 2 .

    Techniques:

    Effects of luteolin on A498 and ACHN cells. A498 and ACHN cells were treated with various concentrations of luteolin for 24 h. Cell viability was determined by MTS reduction assay (a). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (b). * P < 0.05 and ** P < 0.01 versus medium control, n = 4. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of three independent experiments is shown (c). An additional band of HSP90 was indicated by arrow.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Induction of Apoptosis by Luteolin Involving Akt Inactivation in Human 786-O Renal Cell Carcinoma Cells

    doi: 10.1155/2013/109105

    Figure Lengend Snippet: Effects of luteolin on A498 and ACHN cells. A498 and ACHN cells were treated with various concentrations of luteolin for 24 h. Cell viability was determined by MTS reduction assay (a). Protein extracts were isolated and subjected to fluorogenic caspase-3 assay. The intensity of fluorescent signals was expressed as arbitrary unit (b). * P < 0.05 and ** P < 0.01 versus medium control, n = 4. Protein extracts were isolated and subjected to Western blot analysis with indicated antibodies. One of three independent experiments is shown (c). An additional band of HSP90 was indicated by arrow.

    Article Snippet: Human RCC cell lines, 786-O (ATCC CRL1932), A498 (ATCC HTB-44), and ACHN (ATCC CRL-1611) were cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μ g/mL streptomycin and were maintained in a humidified incubator with 5% CO 2 .

    Techniques: Isolation, Caspase-3 Assay, Western Blot