abs against mcl1 Search Results


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  • 99
    Millipore primary antibodies against mcl 1
    Docetaxel sensitizes low <t>MCL-1-expressing</t> cells to BCL-xL inhibition by enhancing mitochondrial priming. a 60 μM of PUMA peptide-induced depolarization in A549 cells treated with chemotherapy drugs at their IC50 levels. A red line is placed at the 40% level to show degree of sensitivity from each treatment. b 60 μM of BAD peptide-induced depolarization in A549 cells treated with 4 nM docetaxel. c Docetaxel sensitizes A549 cells to ABT-263 treatment. A549 cells were treated with indicated chemotherapy drugs at their IC50 levels +/- ABT-263 for 72 h and flow cytometry was applied to measure cell apoptosis. There were three samples in each group. d Docetaxel enhances sensitivity of H727 cells to PUMA and BAD peptide-induced responses. H727 cells were treated with 10 nM docetaxel for 72 h and PUMA and BAD-induced responses were measured by BH3 profiling. A red line is placed at the 40% level to show degree of sensitivity from each treatment. e Docetaxel synergizes with ABT-263 on H727 cell apoptosis. H727 cells were treated with 10 nM docetaxel +/- ABT-263 for 72 h and flow cytometry was performed to measure cell apoptosis. There were three samples in each group. f Expression analysis in A549 and H727 cells after treatment with 25 nM siRNA oligos for 48 h followed by the indicated drugs for another 24 h. g Western blot analysis on PUMA expression in parental and CRISPR-Cas9 engineered A549 cells. h Cell apoptosis analysis in A549 and H727 cells treated with 25 nM siRNA oligos followed by docetaxel and ABT-263 for 72 h. There were three samples in each group. i Cell apoptosis analysis in the parental A549 and CRISPR-Cas9 engineered A549 cells treated with docetaxel and ABT-263 for 72 h. There were three samples in each group. j Expression analysis in A549 cells treated with 25 nM siRNA for 48 h followed by treatment with 4 nM docetaxel for another 24 h. k Synergistic killing of A549 cells from docetaxel combined with siBCL-xL or A-155463. A549 cells were treated with 25 nM siRNA for 48 h followed by 4 nM docetaxel treatment for 72 h (left). A549 cells were treated with 4 nM docetaxel and 100 nM A-1155463 for 72 h (right). There were three samples in each group. l No synergistic killing of A549 cells from docetaxel and ABT-199 combination treatment. A549 cells were treated with 4 nM docetaxel and 1 μM ABT-199 for 72 h. There were three samples in each group
    Primary Antibodies Against Mcl 1, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary antibodies against mcl 1/product/Millipore
    Average 99 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    primary antibodies against mcl 1 - by Bioz Stars, 2020-09
    99/100 stars
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    92
    Santa Cruz Biotechnology antibodies against mcl1
    USP13 genetic deletion suppresses tumor growth. a USP13 depletion in SW-1573 and TOV-21G cells using the CRISPR/Cas9 system. The endogenous USP13 and <t>MCL1</t> protein levels were measured by western blotting. b Cell viability upon USP13 depletion was measured using Cell Counting Kit-8 under hypoxic or oxidative stresses, and error bars indicated standard deviation (each condition contained three biological replicates). c Cell apoptosis upon USP13 depletion was measured using Caspase-Glo 3/7 Assay under hypoxic or oxidative stresses, and error bars indicated standard deviation (each condition contained three biological replicates). d Tumor growth curves and representative tumor images were shown for USP13 -depleted SW-1573 cells subcutaneously implanted into nude mice (scale bar, 10 mm). Error bars indicated standard deviation (10 mice per group). e Tumor growth curves and representative tumor images were shown for USP13 -depleted TOV-21G cells subcutaneously implanted into nude mice (scale bar=10 mm). Error bars indicated standard deviation (10 mice per group). f Tumor growth curves and representative tumor images were shown for USP13 -depleted/ MCL1 -overexpressed SW-1573 cells subcutaneously implanted into nude mice (scale bar, 10 mm). Error bars indicated standard deviation (10 mice per group)
    Antibodies Against Mcl1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibodies against mcl1/product/Santa Cruz Biotechnology
    Average 92 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    antibodies against mcl1 - by Bioz Stars, 2020-09
    92/100 stars
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    99
    Cell Signaling Technology Inc abs against mcl1
    USP13 genetic deletion suppresses tumor growth. a USP13 depletion in SW-1573 and TOV-21G cells using the CRISPR/Cas9 system. The endogenous USP13 and <t>MCL1</t> protein levels were measured by western blotting. b Cell viability upon USP13 depletion was measured using Cell Counting Kit-8 under hypoxic or oxidative stresses, and error bars indicated standard deviation (each condition contained three biological replicates). c Cell apoptosis upon USP13 depletion was measured using Caspase-Glo 3/7 Assay under hypoxic or oxidative stresses, and error bars indicated standard deviation (each condition contained three biological replicates). d Tumor growth curves and representative tumor images were shown for USP13 -depleted SW-1573 cells subcutaneously implanted into nude mice (scale bar, 10 mm). Error bars indicated standard deviation (10 mice per group). e Tumor growth curves and representative tumor images were shown for USP13 -depleted TOV-21G cells subcutaneously implanted into nude mice (scale bar=10 mm). Error bars indicated standard deviation (10 mice per group). f Tumor growth curves and representative tumor images were shown for USP13 -depleted/ MCL1 -overexpressed SW-1573 cells subcutaneously implanted into nude mice (scale bar, 10 mm). Error bars indicated standard deviation (10 mice per group)
    Abs Against Mcl1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/abs against mcl1/product/Cell Signaling Technology Inc
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    abs against mcl1 - by Bioz Stars, 2020-09
    99/100 stars
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    94
    Abcam rat monoclonal antibodies against mcl 1
    SiDcR3 sensitizes hepatocellular carcinoma cells through Bcl-2 family members. Notes: ( A ) The prosurvival Bcl-2 family members Bcl-2, Bcl-xL, IAP-2, survivin, and <t>Mcl-1</t> and proapoptosis member Bax were examined using Western blot analysis after treatment with TRAIL (100 ng/mL) for 24 h. GAPDH was used as loading control. ( B – G ) The ratio of each protein to GAPHD. The differences among different proteins were significant ( P
    Rat Monoclonal Antibodies Against Mcl 1, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat monoclonal antibodies against mcl 1/product/Abcam
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rat monoclonal antibodies against mcl 1 - by Bioz Stars, 2020-09
    94/100 stars
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    Image Search Results


    Docetaxel sensitizes low MCL-1-expressing cells to BCL-xL inhibition by enhancing mitochondrial priming. a 60 μM of PUMA peptide-induced depolarization in A549 cells treated with chemotherapy drugs at their IC50 levels. A red line is placed at the 40% level to show degree of sensitivity from each treatment. b 60 μM of BAD peptide-induced depolarization in A549 cells treated with 4 nM docetaxel. c Docetaxel sensitizes A549 cells to ABT-263 treatment. A549 cells were treated with indicated chemotherapy drugs at their IC50 levels +/- ABT-263 for 72 h and flow cytometry was applied to measure cell apoptosis. There were three samples in each group. d Docetaxel enhances sensitivity of H727 cells to PUMA and BAD peptide-induced responses. H727 cells were treated with 10 nM docetaxel for 72 h and PUMA and BAD-induced responses were measured by BH3 profiling. A red line is placed at the 40% level to show degree of sensitivity from each treatment. e Docetaxel synergizes with ABT-263 on H727 cell apoptosis. H727 cells were treated with 10 nM docetaxel +/- ABT-263 for 72 h and flow cytometry was performed to measure cell apoptosis. There were three samples in each group. f Expression analysis in A549 and H727 cells after treatment with 25 nM siRNA oligos for 48 h followed by the indicated drugs for another 24 h. g Western blot analysis on PUMA expression in parental and CRISPR-Cas9 engineered A549 cells. h Cell apoptosis analysis in A549 and H727 cells treated with 25 nM siRNA oligos followed by docetaxel and ABT-263 for 72 h. There were three samples in each group. i Cell apoptosis analysis in the parental A549 and CRISPR-Cas9 engineered A549 cells treated with docetaxel and ABT-263 for 72 h. There were three samples in each group. j Expression analysis in A549 cells treated with 25 nM siRNA for 48 h followed by treatment with 4 nM docetaxel for another 24 h. k Synergistic killing of A549 cells from docetaxel combined with siBCL-xL or A-155463. A549 cells were treated with 25 nM siRNA for 48 h followed by 4 nM docetaxel treatment for 72 h (left). A549 cells were treated with 4 nM docetaxel and 100 nM A-1155463 for 72 h (right). There were three samples in each group. l No synergistic killing of A549 cells from docetaxel and ABT-199 combination treatment. A549 cells were treated with 4 nM docetaxel and 1 μM ABT-199 for 72 h. There were three samples in each group

    Journal: Cell Death & Disease

    Article Title: Sensitizing non-small cell lung cancer to BCL-xL-targeted apoptosis

    doi: 10.1038/s41419-018-1040-9

    Figure Lengend Snippet: Docetaxel sensitizes low MCL-1-expressing cells to BCL-xL inhibition by enhancing mitochondrial priming. a 60 μM of PUMA peptide-induced depolarization in A549 cells treated with chemotherapy drugs at their IC50 levels. A red line is placed at the 40% level to show degree of sensitivity from each treatment. b 60 μM of BAD peptide-induced depolarization in A549 cells treated with 4 nM docetaxel. c Docetaxel sensitizes A549 cells to ABT-263 treatment. A549 cells were treated with indicated chemotherapy drugs at their IC50 levels +/- ABT-263 for 72 h and flow cytometry was applied to measure cell apoptosis. There were three samples in each group. d Docetaxel enhances sensitivity of H727 cells to PUMA and BAD peptide-induced responses. H727 cells were treated with 10 nM docetaxel for 72 h and PUMA and BAD-induced responses were measured by BH3 profiling. A red line is placed at the 40% level to show degree of sensitivity from each treatment. e Docetaxel synergizes with ABT-263 on H727 cell apoptosis. H727 cells were treated with 10 nM docetaxel +/- ABT-263 for 72 h and flow cytometry was performed to measure cell apoptosis. There were three samples in each group. f Expression analysis in A549 and H727 cells after treatment with 25 nM siRNA oligos for 48 h followed by the indicated drugs for another 24 h. g Western blot analysis on PUMA expression in parental and CRISPR-Cas9 engineered A549 cells. h Cell apoptosis analysis in A549 and H727 cells treated with 25 nM siRNA oligos followed by docetaxel and ABT-263 for 72 h. There were three samples in each group. i Cell apoptosis analysis in the parental A549 and CRISPR-Cas9 engineered A549 cells treated with docetaxel and ABT-263 for 72 h. There were three samples in each group. j Expression analysis in A549 cells treated with 25 nM siRNA for 48 h followed by treatment with 4 nM docetaxel for another 24 h. k Synergistic killing of A549 cells from docetaxel combined with siBCL-xL or A-155463. A549 cells were treated with 25 nM siRNA for 48 h followed by 4 nM docetaxel treatment for 72 h (left). A549 cells were treated with 4 nM docetaxel and 100 nM A-1155463 for 72 h (right). There were three samples in each group. l No synergistic killing of A549 cells from docetaxel and ABT-199 combination treatment. A549 cells were treated with 4 nM docetaxel and 1 μM ABT-199 for 72 h. There were three samples in each group

    Article Snippet: Briefly, the microarray slides were deparaffinized using xylene, rehydrated in a descending series of ethanol and water, then boiled in 10 mM citrate buffer for 10 min. Endogenous peroxidase was applied in 3% hydrogen peroxide in water for 10 min followed by blocking with 2.5% horse serum for 20 min. Primary antibodies against MCL-1 (1:25, #MABC43, EMD Millipore, Billerica, MA, USA) and BCL-xL (1:300, #2764, Cell Signaling, Beverly, MA, USA) were incubated for 1 h at room temperature.

    Techniques: Expressing, Inhibition, Flow Cytometry, Cytometry, Western Blot, CRISPR

    MCL-1 and BCL-xL are upregulated in a subset of NSCLCs. a Analysis of TCGA database on MCL-1 and BCL- xL gene amplifications in NSCLC patients ( n = 230). amp, amplification. b Representative immunohistochemical staining results of NSCLC tissue. c Summary of immunohistochemical staining results of NSCLC tissue and adjacent normal lung tissue. Red indicates positive immunohistochemical staining, and blue indicates negative immunohistochemical staining

    Journal: Cell Death & Disease

    Article Title: Sensitizing non-small cell lung cancer to BCL-xL-targeted apoptosis

    doi: 10.1038/s41419-018-1040-9

    Figure Lengend Snippet: MCL-1 and BCL-xL are upregulated in a subset of NSCLCs. a Analysis of TCGA database on MCL-1 and BCL- xL gene amplifications in NSCLC patients ( n = 230). amp, amplification. b Representative immunohistochemical staining results of NSCLC tissue. c Summary of immunohistochemical staining results of NSCLC tissue and adjacent normal lung tissue. Red indicates positive immunohistochemical staining, and blue indicates negative immunohistochemical staining

    Article Snippet: Briefly, the microarray slides were deparaffinized using xylene, rehydrated in a descending series of ethanol and water, then boiled in 10 mM citrate buffer for 10 min. Endogenous peroxidase was applied in 3% hydrogen peroxide in water for 10 min followed by blocking with 2.5% horse serum for 20 min. Primary antibodies against MCL-1 (1:25, #MABC43, EMD Millipore, Billerica, MA, USA) and BCL-xL (1:300, #2764, Cell Signaling, Beverly, MA, USA) were incubated for 1 h at room temperature.

    Techniques: Amplification, Immunohistochemistry, Staining

    Variable apoptotic response from NSCLS cells to MCL-1 and BCL-xL inhibition. a Expression profile of BCL-2 family proteins in NSCLC cells. b Knockdown of MCL-1 expression in NSCLC cells 48 h after transfection with 25 nM siRNA oligos. siSCR: scramble siRNA; siMCL-1a: MCL-1a siRNA. c The percentage of apoptotic cells after siMCL-1a and AT-263 treatment. Cells were transfected with 25 nM siMCL-1a for 48 h and then treated with 1 μM ABT-263 for 36 h. Cell apoptosis was measured with flow cytometry. There were 3 samples in each group. d The percentage of apoptotic cells after siMCL-1a and ABT-199 treatment. Cells were transfected with 25 nM siMCL-1a or siSCR for 48 h and then treated with 1 μM ABT-199 for 36 h. Cell apoptosis was measured with flow cytometry. There were three samples in each group. e The percentage of apoptotic cells after siMCL-1a and siBCL-xL treatment. Cells were transfected with 25 nM siRNA oligos for 72 h and then analyzed for cell apoptosis with flow cytometry. There were three samples in each group. f The percentage of apoptotic cells after siMCL-1a and A-1155463 treatment. Cells were transfected with 25 nM siRNA oligos for 48 h then treated with 100 nM A-1155463 for 36 h. Cell apoptosis was measured with flow cytometry. There were three samples in each group

    Journal: Cell Death & Disease

    Article Title: Sensitizing non-small cell lung cancer to BCL-xL-targeted apoptosis

    doi: 10.1038/s41419-018-1040-9

    Figure Lengend Snippet: Variable apoptotic response from NSCLS cells to MCL-1 and BCL-xL inhibition. a Expression profile of BCL-2 family proteins in NSCLC cells. b Knockdown of MCL-1 expression in NSCLC cells 48 h after transfection with 25 nM siRNA oligos. siSCR: scramble siRNA; siMCL-1a: MCL-1a siRNA. c The percentage of apoptotic cells after siMCL-1a and AT-263 treatment. Cells were transfected with 25 nM siMCL-1a for 48 h and then treated with 1 μM ABT-263 for 36 h. Cell apoptosis was measured with flow cytometry. There were 3 samples in each group. d The percentage of apoptotic cells after siMCL-1a and ABT-199 treatment. Cells were transfected with 25 nM siMCL-1a or siSCR for 48 h and then treated with 1 μM ABT-199 for 36 h. Cell apoptosis was measured with flow cytometry. There were three samples in each group. e The percentage of apoptotic cells after siMCL-1a and siBCL-xL treatment. Cells were transfected with 25 nM siRNA oligos for 72 h and then analyzed for cell apoptosis with flow cytometry. There were three samples in each group. f The percentage of apoptotic cells after siMCL-1a and A-1155463 treatment. Cells were transfected with 25 nM siRNA oligos for 48 h then treated with 100 nM A-1155463 for 36 h. Cell apoptosis was measured with flow cytometry. There were three samples in each group

    Article Snippet: Briefly, the microarray slides were deparaffinized using xylene, rehydrated in a descending series of ethanol and water, then boiled in 10 mM citrate buffer for 10 min. Endogenous peroxidase was applied in 3% hydrogen peroxide in water for 10 min followed by blocking with 2.5% horse serum for 20 min. Primary antibodies against MCL-1 (1:25, #MABC43, EMD Millipore, Billerica, MA, USA) and BCL-xL (1:300, #2764, Cell Signaling, Beverly, MA, USA) were incubated for 1 h at room temperature.

    Techniques: Inhibition, Expressing, Transfection, Flow Cytometry, Cytometry

    Sensitivity to MCL-1 and BCL-xL inhibition positively correlates with mitochondrial priming. a Binding affinity of PUMA, BAD, and NOXA-derived peptides to the BCL-2 family proteins. b BH3 profiling of 10 NSCLC cell lines using indicated peptides (60 μM of each peptide). The mean responses induced by the PUMA, BAD, or NOXA-derived peptides are shown. A red line is placed at the 40% level to show degree of sensitivity from each treatment. There were three samples in each group. ND, not detected. c Correlation between siMCL-1 and ABT-263 induced apoptosis and the PUMA peptide-induced depolarization. d Correlation between BAD and NOXA peptide-induced depolarization and the PUMA peptide-induced depolarization. e Correlation between MCL-1 expression level and the PUMA peptide-induced depolarization

    Journal: Cell Death & Disease

    Article Title: Sensitizing non-small cell lung cancer to BCL-xL-targeted apoptosis

    doi: 10.1038/s41419-018-1040-9

    Figure Lengend Snippet: Sensitivity to MCL-1 and BCL-xL inhibition positively correlates with mitochondrial priming. a Binding affinity of PUMA, BAD, and NOXA-derived peptides to the BCL-2 family proteins. b BH3 profiling of 10 NSCLC cell lines using indicated peptides (60 μM of each peptide). The mean responses induced by the PUMA, BAD, or NOXA-derived peptides are shown. A red line is placed at the 40% level to show degree of sensitivity from each treatment. There were three samples in each group. ND, not detected. c Correlation between siMCL-1 and ABT-263 induced apoptosis and the PUMA peptide-induced depolarization. d Correlation between BAD and NOXA peptide-induced depolarization and the PUMA peptide-induced depolarization. e Correlation between MCL-1 expression level and the PUMA peptide-induced depolarization

    Article Snippet: Briefly, the microarray slides were deparaffinized using xylene, rehydrated in a descending series of ethanol and water, then boiled in 10 mM citrate buffer for 10 min. Endogenous peroxidase was applied in 3% hydrogen peroxide in water for 10 min followed by blocking with 2.5% horse serum for 20 min. Primary antibodies against MCL-1 (1:25, #MABC43, EMD Millipore, Billerica, MA, USA) and BCL-xL (1:300, #2764, Cell Signaling, Beverly, MA, USA) were incubated for 1 h at room temperature.

    Techniques: Inhibition, Binding Assay, Derivative Assay, Expressing

    MCL-1 expression level correlates with sensitivity of NSCLC to MCL-1 and BCL-xL inhibition. Expression levels of BCL-2, BCL-xL, BIM, BID, PUMA, and MCL-1 were quantified using the Image Studio Ver 5.0 software. The level for each protein in SK-LU-1 cells was set as 1, and relative expression levels in other cells was calculated by comparing to SK-LU-1 cells. a – f The percentages of apoptotic cells induced by knockdown of MCL-1 combined with ABT-263 were plotted against relative expression levels of individual proteins in the NSCLC cells. The non-parametric one-tailed Spearman’s correlation test was applied to perform statistical analysis

    Journal: Cell Death & Disease

    Article Title: Sensitizing non-small cell lung cancer to BCL-xL-targeted apoptosis

    doi: 10.1038/s41419-018-1040-9

    Figure Lengend Snippet: MCL-1 expression level correlates with sensitivity of NSCLC to MCL-1 and BCL-xL inhibition. Expression levels of BCL-2, BCL-xL, BIM, BID, PUMA, and MCL-1 were quantified using the Image Studio Ver 5.0 software. The level for each protein in SK-LU-1 cells was set as 1, and relative expression levels in other cells was calculated by comparing to SK-LU-1 cells. a – f The percentages of apoptotic cells induced by knockdown of MCL-1 combined with ABT-263 were plotted against relative expression levels of individual proteins in the NSCLC cells. The non-parametric one-tailed Spearman’s correlation test was applied to perform statistical analysis

    Article Snippet: Briefly, the microarray slides were deparaffinized using xylene, rehydrated in a descending series of ethanol and water, then boiled in 10 mM citrate buffer for 10 min. Endogenous peroxidase was applied in 3% hydrogen peroxide in water for 10 min followed by blocking with 2.5% horse serum for 20 min. Primary antibodies against MCL-1 (1:25, #MABC43, EMD Millipore, Billerica, MA, USA) and BCL-xL (1:300, #2764, Cell Signaling, Beverly, MA, USA) were incubated for 1 h at room temperature.

    Techniques: Expressing, Inhibition, Software, One-tailed Test

    Mcl-1 promotes cell migration in lung cancer cells. ( a ) Western blot showing different expression levels of Mcl-1 and Bcl-x L in three different NSCLC cell lines. ( b ) Western blot of H1299 and H460 cells transfected with control or Mcl-1 siRNA showing efficient knockdown of Mcl-1 in both cell lines. ( c ) Representative images of wound-healing assay showing control and Mcl-1 knockdown A549 cells at 0 and 24 h post scratch. ( d ) Summary bar graphs of the percentage closure in 24 h of control and Mcl-1 knockdown A549, H1299 and H460 cells (mean±S.E.; *, P

    Journal: Cell Death & Disease

    Article Title: Mcl-1 promotes lung cancer cell migration by directly interacting with VDAC to increase mitochondrial Ca2+ uptake and reactive oxygen species generation

    doi: 10.1038/cddis.2014.419

    Figure Lengend Snippet: Mcl-1 promotes cell migration in lung cancer cells. ( a ) Western blot showing different expression levels of Mcl-1 and Bcl-x L in three different NSCLC cell lines. ( b ) Western blot of H1299 and H460 cells transfected with control or Mcl-1 siRNA showing efficient knockdown of Mcl-1 in both cell lines. ( c ) Representative images of wound-healing assay showing control and Mcl-1 knockdown A549 cells at 0 and 24 h post scratch. ( d ) Summary bar graphs of the percentage closure in 24 h of control and Mcl-1 knockdown A549, H1299 and H460 cells (mean±S.E.; *, P

    Article Snippet: Western blot was performed using antibodies against Mcl-1 (EMD Millipore Billerica, MA, USA), Bcl-xL (BD Biosciences, San Jose, CA, USA), GST (ViroGen Corporation, Watertown, MA, USA) and VDAC1/3 (Abcam; Cambridge, MA, USA).

    Techniques: Migration, Western Blot, Expressing, Transfection, Wound Healing Assay

    Mcl-1 does not affect cell proliferation in lung cancer cells. ( a ) Western blot showing efficient knockdown of Mcl-1 by shRNA-transfected stable cell lines from A549, H1299 and H460 cells. Bcl-x L and VDAC levels are unaffected. ( b ) Summary of proliferation in control Mcl-1 WT and Mcl-1 knockdown lung cancer cells (mean±S.E.; P > 0.05; student's t -test)

    Journal: Cell Death & Disease

    Article Title: Mcl-1 promotes lung cancer cell migration by directly interacting with VDAC to increase mitochondrial Ca2+ uptake and reactive oxygen species generation

    doi: 10.1038/cddis.2014.419

    Figure Lengend Snippet: Mcl-1 does not affect cell proliferation in lung cancer cells. ( a ) Western blot showing efficient knockdown of Mcl-1 by shRNA-transfected stable cell lines from A549, H1299 and H460 cells. Bcl-x L and VDAC levels are unaffected. ( b ) Summary of proliferation in control Mcl-1 WT and Mcl-1 knockdown lung cancer cells (mean±S.E.; P > 0.05; student's t -test)

    Article Snippet: Western blot was performed using antibodies against Mcl-1 (EMD Millipore Billerica, MA, USA), Bcl-xL (BD Biosciences, San Jose, CA, USA), GST (ViroGen Corporation, Watertown, MA, USA) and VDAC1/3 (Abcam; Cambridge, MA, USA).

    Techniques: Western Blot, shRNA, Transfection, Stable Transfection

    Mcl-1 and VDAC1/3 interact in vivo to increase [Ca 2+ ] mito uptake. ( a ) An in situ proximity ligation assay (PLA) showing the interaction between MCL-1 and VDAC1 and 3 (red) in A549 cells counterstained with DAPI (blue). ( b ) Quantification (mean±S.E.) of the PLA signal normalized to cell number (* P

    Journal: Cell Death & Disease

    Article Title: Mcl-1 promotes lung cancer cell migration by directly interacting with VDAC to increase mitochondrial Ca2+ uptake and reactive oxygen species generation

    doi: 10.1038/cddis.2014.419

    Figure Lengend Snippet: Mcl-1 and VDAC1/3 interact in vivo to increase [Ca 2+ ] mito uptake. ( a ) An in situ proximity ligation assay (PLA) showing the interaction between MCL-1 and VDAC1 and 3 (red) in A549 cells counterstained with DAPI (blue). ( b ) Quantification (mean±S.E.) of the PLA signal normalized to cell number (* P

    Article Snippet: Western blot was performed using antibodies against Mcl-1 (EMD Millipore Billerica, MA, USA), Bcl-xL (BD Biosciences, San Jose, CA, USA), GST (ViroGen Corporation, Watertown, MA, USA) and VDAC1/3 (Abcam; Cambridge, MA, USA).

    Techniques: In Vivo, In Situ, Proximity Ligation Assay

    Mcl-1 increases reactive oxygen species (ROS) production in A549 cancer cells by increasing [Ca 2+ ] mito . ( a ) Representative images showing fluorescence signal from A549 cells before and after loading with the ROS indicator dihydrorhodamine 123. ( b ) Summary bar graphs showing the mean±S.E. (300–400 cells, pooled from three independent experiments; * P

    Journal: Cell Death & Disease

    Article Title: Mcl-1 promotes lung cancer cell migration by directly interacting with VDAC to increase mitochondrial Ca2+ uptake and reactive oxygen species generation

    doi: 10.1038/cddis.2014.419

    Figure Lengend Snippet: Mcl-1 increases reactive oxygen species (ROS) production in A549 cancer cells by increasing [Ca 2+ ] mito . ( a ) Representative images showing fluorescence signal from A549 cells before and after loading with the ROS indicator dihydrorhodamine 123. ( b ) Summary bar graphs showing the mean±S.E. (300–400 cells, pooled from three independent experiments; * P

    Article Snippet: Western blot was performed using antibodies against Mcl-1 (EMD Millipore Billerica, MA, USA), Bcl-xL (BD Biosciences, San Jose, CA, USA), GST (ViroGen Corporation, Watertown, MA, USA) and VDAC1/3 (Abcam; Cambridge, MA, USA).

    Techniques: Fluorescence

    Mcl-1 binds strongly to VDAC1 and 3 and binds to VDAC1 with greater apparent affinity than Bcl-x L . ( a ) Human Mcl-1 expressed in MEF cells and detected by western blot after pulldown by GST-fusion proteins of VDAC1, 2 or 3 is shown in the upper lanes with loading control blots of GST depicted below. ( b ) GST-fusion proteins of VDAC1, 2 or 3 detected by anti-GST antibody after pulldown by purified His-tagged Mcl-1 is shown in the upper lanes with the loading control blots of Mcl-1 depicted below. Lanes labeled BSA are negative controls showing GST-VDACs are not pulled down by beads alone. ( c ) Western blot detection of Mcl-1 and Bcl-x L pulled down by GST-VDAC1 from MEF cell lysates pretreated with control or VDAC-based peptides (N-ter and L14-15). ( d ) Western blot of recombinant purified Mcl-1 and Bcl-x L (0.2 μ g) pulled down by GST-VDAC1. All blots are representative of at least two independent experiments

    Journal: Cell Death & Disease

    Article Title: Mcl-1 promotes lung cancer cell migration by directly interacting with VDAC to increase mitochondrial Ca2+ uptake and reactive oxygen species generation

    doi: 10.1038/cddis.2014.419

    Figure Lengend Snippet: Mcl-1 binds strongly to VDAC1 and 3 and binds to VDAC1 with greater apparent affinity than Bcl-x L . ( a ) Human Mcl-1 expressed in MEF cells and detected by western blot after pulldown by GST-fusion proteins of VDAC1, 2 or 3 is shown in the upper lanes with loading control blots of GST depicted below. ( b ) GST-fusion proteins of VDAC1, 2 or 3 detected by anti-GST antibody after pulldown by purified His-tagged Mcl-1 is shown in the upper lanes with the loading control blots of Mcl-1 depicted below. Lanes labeled BSA are negative controls showing GST-VDACs are not pulled down by beads alone. ( c ) Western blot detection of Mcl-1 and Bcl-x L pulled down by GST-VDAC1 from MEF cell lysates pretreated with control or VDAC-based peptides (N-ter and L14-15). ( d ) Western blot of recombinant purified Mcl-1 and Bcl-x L (0.2 μ g) pulled down by GST-VDAC1. All blots are representative of at least two independent experiments

    Article Snippet: Western blot was performed using antibodies against Mcl-1 (EMD Millipore Billerica, MA, USA), Bcl-xL (BD Biosciences, San Jose, CA, USA), GST (ViroGen Corporation, Watertown, MA, USA) and VDAC1/3 (Abcam; Cambridge, MA, USA).

    Techniques: Western Blot, Purification, Labeling, Recombinant

    Mcl-1 promotes cell migration by a ROS-dependent mechanism. ( a ) Representative images of the migratory tracks created by both WT and Mcl-1 knockdown A549, H1299 and H460 cells on colloidal gold-coated coverslips in a 24-h time period. ( b , c ) The effect of Mcl-1 knockdown and Mcl-1/VDAC inhibition on ROS levels and migration. Bar graphs showing the effect of VDAC-based peptides (N-ter and L14-15; 2 μ M) on ROS levels assessed by RH123 ( b ), and cell migration assessed by the migratory track area ( c ) on the cell lines indicated. Data are presented as mean±S.E. normalized to values obtained under control conditions, and represent 100–200 cells pooled from multiple coverslips run on at least two independent days ( P

    Journal: Cell Death & Disease

    Article Title: Mcl-1 promotes lung cancer cell migration by directly interacting with VDAC to increase mitochondrial Ca2+ uptake and reactive oxygen species generation

    doi: 10.1038/cddis.2014.419

    Figure Lengend Snippet: Mcl-1 promotes cell migration by a ROS-dependent mechanism. ( a ) Representative images of the migratory tracks created by both WT and Mcl-1 knockdown A549, H1299 and H460 cells on colloidal gold-coated coverslips in a 24-h time period. ( b , c ) The effect of Mcl-1 knockdown and Mcl-1/VDAC inhibition on ROS levels and migration. Bar graphs showing the effect of VDAC-based peptides (N-ter and L14-15; 2 μ M) on ROS levels assessed by RH123 ( b ), and cell migration assessed by the migratory track area ( c ) on the cell lines indicated. Data are presented as mean±S.E. normalized to values obtained under control conditions, and represent 100–200 cells pooled from multiple coverslips run on at least two independent days ( P

    Article Snippet: Western blot was performed using antibodies against Mcl-1 (EMD Millipore Billerica, MA, USA), Bcl-xL (BD Biosciences, San Jose, CA, USA), GST (ViroGen Corporation, Watertown, MA, USA) and VDAC1/3 (Abcam; Cambridge, MA, USA).

    Techniques: Migration, Inhibition

    USP13 genetic deletion suppresses tumor growth. a USP13 depletion in SW-1573 and TOV-21G cells using the CRISPR/Cas9 system. The endogenous USP13 and MCL1 protein levels were measured by western blotting. b Cell viability upon USP13 depletion was measured using Cell Counting Kit-8 under hypoxic or oxidative stresses, and error bars indicated standard deviation (each condition contained three biological replicates). c Cell apoptosis upon USP13 depletion was measured using Caspase-Glo 3/7 Assay under hypoxic or oxidative stresses, and error bars indicated standard deviation (each condition contained three biological replicates). d Tumor growth curves and representative tumor images were shown for USP13 -depleted SW-1573 cells subcutaneously implanted into nude mice (scale bar, 10 mm). Error bars indicated standard deviation (10 mice per group). e Tumor growth curves and representative tumor images were shown for USP13 -depleted TOV-21G cells subcutaneously implanted into nude mice (scale bar=10 mm). Error bars indicated standard deviation (10 mice per group). f Tumor growth curves and representative tumor images were shown for USP13 -depleted/ MCL1 -overexpressed SW-1573 cells subcutaneously implanted into nude mice (scale bar, 10 mm). Error bars indicated standard deviation (10 mice per group)

    Journal: Nature Communications

    Article Title: Deubiquitinase USP13 dictates MCL1 stability and sensitivity to BH3 mimetic inhibitors

    doi: 10.1038/s41467-017-02693-9

    Figure Lengend Snippet: USP13 genetic deletion suppresses tumor growth. a USP13 depletion in SW-1573 and TOV-21G cells using the CRISPR/Cas9 system. The endogenous USP13 and MCL1 protein levels were measured by western blotting. b Cell viability upon USP13 depletion was measured using Cell Counting Kit-8 under hypoxic or oxidative stresses, and error bars indicated standard deviation (each condition contained three biological replicates). c Cell apoptosis upon USP13 depletion was measured using Caspase-Glo 3/7 Assay under hypoxic or oxidative stresses, and error bars indicated standard deviation (each condition contained three biological replicates). d Tumor growth curves and representative tumor images were shown for USP13 -depleted SW-1573 cells subcutaneously implanted into nude mice (scale bar, 10 mm). Error bars indicated standard deviation (10 mice per group). e Tumor growth curves and representative tumor images were shown for USP13 -depleted TOV-21G cells subcutaneously implanted into nude mice (scale bar=10 mm). Error bars indicated standard deviation (10 mice per group). f Tumor growth curves and representative tumor images were shown for USP13 -depleted/ MCL1 -overexpressed SW-1573 cells subcutaneously implanted into nude mice (scale bar, 10 mm). Error bars indicated standard deviation (10 mice per group)

    Article Snippet: The tissue slides were deparaffinized, treated with 3% H2 O2 for 10 min, autoclaved in 10 mM citric sodium (pH 6.0) for 30 min to unmask antigens, rinsed in PBS, and then incubated with the primary antibodies against MCL1 (Santa Cruz, sc-819, 1:100) or USP13 (Santa Cruz, sc-514416, 1:250) at 4 °C overnight, followed by incubation with biotinylated secondary antibody for 1 hour at room temperature.

    Techniques: CRISPR, Western Blot, Cell Counting, Standard Deviation, Caspase-Glo Assay, Mouse Assay

    Targeting USP13 sensitizes tumor cells to BH3 mimetics. a USP13 or MCL1 was knocked out in SW-1573 and TOV-21G cells using CRISPR/Cas9, and cell viability in the presence of ABT-263 was analyzed by Cell Counting Kit-8. Error bars indicated standard deviation (each condition contained three biological replicates). b SW-1573 and TOV-21G cells were treated with spautin-1 (10 μM) and MG132 (10 μM) for 6 h. Cell lysates were analyzed by western blotting using antibodies against USP13, MCL1 or H3. c Cells were treated with spautin-1 at indicated concentrations or for different time frames, and cell lysates were analyzed by western blotting using antibodies against USP13, MCL1 or H3. d Examples of synergistic activities of spautin-1 and ABT-263 in SW-1573 and TOV-21G cells. Left: heatmaps of growth inhibition; right: heatmaps of Bliss synergy scores. e Cells were treated with DMSO, spautin-1 (1 μM) and/or ABT-263 (1 μM). The remaining cells were stained with crystal violet and quantified (scale bar=3.5 mm). Error bars indicated standard deviation (each condition contained three biological replicates)

    Journal: Nature Communications

    Article Title: Deubiquitinase USP13 dictates MCL1 stability and sensitivity to BH3 mimetic inhibitors

    doi: 10.1038/s41467-017-02693-9

    Figure Lengend Snippet: Targeting USP13 sensitizes tumor cells to BH3 mimetics. a USP13 or MCL1 was knocked out in SW-1573 and TOV-21G cells using CRISPR/Cas9, and cell viability in the presence of ABT-263 was analyzed by Cell Counting Kit-8. Error bars indicated standard deviation (each condition contained three biological replicates). b SW-1573 and TOV-21G cells were treated with spautin-1 (10 μM) and MG132 (10 μM) for 6 h. Cell lysates were analyzed by western blotting using antibodies against USP13, MCL1 or H3. c Cells were treated with spautin-1 at indicated concentrations or for different time frames, and cell lysates were analyzed by western blotting using antibodies against USP13, MCL1 or H3. d Examples of synergistic activities of spautin-1 and ABT-263 in SW-1573 and TOV-21G cells. Left: heatmaps of growth inhibition; right: heatmaps of Bliss synergy scores. e Cells were treated with DMSO, spautin-1 (1 μM) and/or ABT-263 (1 μM). The remaining cells were stained with crystal violet and quantified (scale bar=3.5 mm). Error bars indicated standard deviation (each condition contained three biological replicates)

    Article Snippet: The tissue slides were deparaffinized, treated with 3% H2 O2 for 10 min, autoclaved in 10 mM citric sodium (pH 6.0) for 30 min to unmask antigens, rinsed in PBS, and then incubated with the primary antibodies against MCL1 (Santa Cruz, sc-819, 1:100) or USP13 (Santa Cruz, sc-514416, 1:250) at 4 °C overnight, followed by incubation with biotinylated secondary antibody for 1 hour at room temperature.

    Techniques: CRISPR, Cell Counting, Standard Deviation, Western Blot, Inhibition, Staining

    Identification of USP13 as a candidate MCL1 deubiquitinase. a The quantification of MCL1 protein levels upon knockdown of each DUB in HEK293T cells. 84 siRNA pools were individually transfected into HEK293T cells, endogenous MCL1 was detected by western blotting and protein bands were quantified using the Image J software. b Classification of the 84 human DUBs included in siRNA screen. c Dot plot of copy number alterations for 20 candidate MCL DUB genes in TCGA cancers. d Validation of the candidate MCL1 DUBs in HEK293T cells. Four different siRNAs targeting USP9X , OTUB2 , or USP13 were transfected into HEK293T cells, and endogenous expression levels of indicated proteins were measured by western blot analysis

    Journal: Nature Communications

    Article Title: Deubiquitinase USP13 dictates MCL1 stability and sensitivity to BH3 mimetic inhibitors

    doi: 10.1038/s41467-017-02693-9

    Figure Lengend Snippet: Identification of USP13 as a candidate MCL1 deubiquitinase. a The quantification of MCL1 protein levels upon knockdown of each DUB in HEK293T cells. 84 siRNA pools were individually transfected into HEK293T cells, endogenous MCL1 was detected by western blotting and protein bands were quantified using the Image J software. b Classification of the 84 human DUBs included in siRNA screen. c Dot plot of copy number alterations for 20 candidate MCL DUB genes in TCGA cancers. d Validation of the candidate MCL1 DUBs in HEK293T cells. Four different siRNAs targeting USP9X , OTUB2 , or USP13 were transfected into HEK293T cells, and endogenous expression levels of indicated proteins were measured by western blot analysis

    Article Snippet: The tissue slides were deparaffinized, treated with 3% H2 O2 for 10 min, autoclaved in 10 mM citric sodium (pH 6.0) for 30 min to unmask antigens, rinsed in PBS, and then incubated with the primary antibodies against MCL1 (Santa Cruz, sc-819, 1:100) or USP13 (Santa Cruz, sc-514416, 1:250) at 4 °C overnight, followed by incubation with biotinylated secondary antibody for 1 hour at room temperature.

    Techniques: Transfection, Western Blot, Software, Expressing

    USP13 and MCL1 are correlatively upregulated in cancer. a Pan-cancer analysis of USP13 and MCL1 genomic alterations in cBioportal database. b Copy number analysis of USP13 and MCL1 in TCGA lung adenocarcinoma, lung squamous cell carcinoma and ovarian serous carcinoma samples. Color scale: amplification in red and deletion in blue. c Expression of USP13 and MCL1 in human lung cancer and ovarian cancer cell lines. We used 22 lung cancer and 33 ovarian cancer cell lines to determine the endogenous protein levels of USP13 and MCL1. d Representative images for immunohistochemical staining of USP13 and MCL1 protein in human ovarian cancer samples, normal lung and lung cancer tissues (scale bar=50 µm). Statistical significance of correlation between USP13 and MCL1 protein levels was determined by a χ 2 (chi-squared) test. R represented the correlation coefficients. e Positive correlation of USP13 with MCL1 protein expression in human ovarian cancer tissues, and enrichment of USP13 or MCL1 expression in human lung cancer specimens relative to normal lung tissues. Forty-five human ovarian cancer samples were categorized into two groups (13 USP13-low and 32 USP13-high), and MCL1 expression status was indicated in each group. Thirty human normal lung or lung cancer tissues were classified based on the USP13 or MCL protein expression status

    Journal: Nature Communications

    Article Title: Deubiquitinase USP13 dictates MCL1 stability and sensitivity to BH3 mimetic inhibitors

    doi: 10.1038/s41467-017-02693-9

    Figure Lengend Snippet: USP13 and MCL1 are correlatively upregulated in cancer. a Pan-cancer analysis of USP13 and MCL1 genomic alterations in cBioportal database. b Copy number analysis of USP13 and MCL1 in TCGA lung adenocarcinoma, lung squamous cell carcinoma and ovarian serous carcinoma samples. Color scale: amplification in red and deletion in blue. c Expression of USP13 and MCL1 in human lung cancer and ovarian cancer cell lines. We used 22 lung cancer and 33 ovarian cancer cell lines to determine the endogenous protein levels of USP13 and MCL1. d Representative images for immunohistochemical staining of USP13 and MCL1 protein in human ovarian cancer samples, normal lung and lung cancer tissues (scale bar=50 µm). Statistical significance of correlation between USP13 and MCL1 protein levels was determined by a χ 2 (chi-squared) test. R represented the correlation coefficients. e Positive correlation of USP13 with MCL1 protein expression in human ovarian cancer tissues, and enrichment of USP13 or MCL1 expression in human lung cancer specimens relative to normal lung tissues. Forty-five human ovarian cancer samples were categorized into two groups (13 USP13-low and 32 USP13-high), and MCL1 expression status was indicated in each group. Thirty human normal lung or lung cancer tissues were classified based on the USP13 or MCL protein expression status

    Article Snippet: The tissue slides were deparaffinized, treated with 3% H2 O2 for 10 min, autoclaved in 10 mM citric sodium (pH 6.0) for 30 min to unmask antigens, rinsed in PBS, and then incubated with the primary antibodies against MCL1 (Santa Cruz, sc-819, 1:100) or USP13 (Santa Cruz, sc-514416, 1:250) at 4 °C overnight, followed by incubation with biotinylated secondary antibody for 1 hour at room temperature.

    Techniques: Amplification, Expressing, Immunohistochemistry, Staining

    USP13 interacts with and deubiquitinates MCL1. a 3 × FLAG-tagged USP13 was transfected into HEK293T cells. Cells were treated with MG-132 for 8 h and USP13 was immunoprecipitated using an anti-FLAG antibody. Co-immunoprecipitated MCL1 was detected using an anti-MCL1 antibody. b Endogenous USP13 and MCL1 were immunoprecipitated from A549, HEY and SW-1573 cells using an anti-USP13 antibody or an anti-MCL1 antibody respectively. Co-immunoprecipitated endogenous MCL1 or USP13 was analyzed by western blotting. c Left: Purified His-MCL1 and His-USP13 proteins were co-incubated and immunoprecipitated with an anti-USP13 antibody. Co-immunoprecipitated MCL1 was detected using an anti-MCL1 antibody. Right: FLAG-USP13 or FLAG-USP13-C345A was transfected into HEK293T cells and immunoprecipitated using an anti-FLAG antibody. Co-immunoprecipitated MCL1 was detected using an anti-MCL1 antibody. d A schematic of full-length USP13 (FL), full-length MCL1 (FL) and their deletion mutants (M1, M2, U1, U2). e USP13 was co-transfected with the MCL1 deletion mutants, and MCL1 was co-transfected with the USP13 deletion mutants. Interactions were analyzed using the co-immunoprecipitation assay. f MCL1 was co-transfected with FLAG-USP13 or the FLAG-USP13-C345A mutant, with or without HA-ubiquitin. The ubiquitinated MCL1 was measured using an in vivo ubiquitination assay. g Purified His-MCL1 and His-USP13 proteins were co-incubated in the ubiquitination system. The ubiquitinated MCL1 was measured using an in vitro ubiquitination assay

    Journal: Nature Communications

    Article Title: Deubiquitinase USP13 dictates MCL1 stability and sensitivity to BH3 mimetic inhibitors

    doi: 10.1038/s41467-017-02693-9

    Figure Lengend Snippet: USP13 interacts with and deubiquitinates MCL1. a 3 × FLAG-tagged USP13 was transfected into HEK293T cells. Cells were treated with MG-132 for 8 h and USP13 was immunoprecipitated using an anti-FLAG antibody. Co-immunoprecipitated MCL1 was detected using an anti-MCL1 antibody. b Endogenous USP13 and MCL1 were immunoprecipitated from A549, HEY and SW-1573 cells using an anti-USP13 antibody or an anti-MCL1 antibody respectively. Co-immunoprecipitated endogenous MCL1 or USP13 was analyzed by western blotting. c Left: Purified His-MCL1 and His-USP13 proteins were co-incubated and immunoprecipitated with an anti-USP13 antibody. Co-immunoprecipitated MCL1 was detected using an anti-MCL1 antibody. Right: FLAG-USP13 or FLAG-USP13-C345A was transfected into HEK293T cells and immunoprecipitated using an anti-FLAG antibody. Co-immunoprecipitated MCL1 was detected using an anti-MCL1 antibody. d A schematic of full-length USP13 (FL), full-length MCL1 (FL) and their deletion mutants (M1, M2, U1, U2). e USP13 was co-transfected with the MCL1 deletion mutants, and MCL1 was co-transfected with the USP13 deletion mutants. Interactions were analyzed using the co-immunoprecipitation assay. f MCL1 was co-transfected with FLAG-USP13 or the FLAG-USP13-C345A mutant, with or without HA-ubiquitin. The ubiquitinated MCL1 was measured using an in vivo ubiquitination assay. g Purified His-MCL1 and His-USP13 proteins were co-incubated in the ubiquitination system. The ubiquitinated MCL1 was measured using an in vitro ubiquitination assay

    Article Snippet: The tissue slides were deparaffinized, treated with 3% H2 O2 for 10 min, autoclaved in 10 mM citric sodium (pH 6.0) for 30 min to unmask antigens, rinsed in PBS, and then incubated with the primary antibodies against MCL1 (Santa Cruz, sc-819, 1:100) or USP13 (Santa Cruz, sc-514416, 1:250) at 4 °C overnight, followed by incubation with biotinylated secondary antibody for 1 hour at room temperature.

    Techniques: Transfection, Immunoprecipitation, Western Blot, Purification, Incubation, Co-Immunoprecipitation Assay, Mutagenesis, In Vivo, Ubiquitin Assay, In Vitro

    USP13 promotes MCL1 protein stability. a USP13 knockdown by siRNAs decreased MCL1 protein levels in human ovarian cancer (TOV-21G, HEY and OVCA433) and lung cancer (SW-1573, NCI-H441 and A549) cell lines. b Quantitative PCR analysis of USP13 and MCL1 mRNA levels in TOV-21G, SW-1573 and OVCA433 cells upon USP13 knockdown (* P

    Journal: Nature Communications

    Article Title: Deubiquitinase USP13 dictates MCL1 stability and sensitivity to BH3 mimetic inhibitors

    doi: 10.1038/s41467-017-02693-9

    Figure Lengend Snippet: USP13 promotes MCL1 protein stability. a USP13 knockdown by siRNAs decreased MCL1 protein levels in human ovarian cancer (TOV-21G, HEY and OVCA433) and lung cancer (SW-1573, NCI-H441 and A549) cell lines. b Quantitative PCR analysis of USP13 and MCL1 mRNA levels in TOV-21G, SW-1573 and OVCA433 cells upon USP13 knockdown (* P

    Article Snippet: The tissue slides were deparaffinized, treated with 3% H2 O2 for 10 min, autoclaved in 10 mM citric sodium (pH 6.0) for 30 min to unmask antigens, rinsed in PBS, and then incubated with the primary antibodies against MCL1 (Santa Cruz, sc-819, 1:100) or USP13 (Santa Cruz, sc-514416, 1:250) at 4 °C overnight, followed by incubation with biotinylated secondary antibody for 1 hour at room temperature.

    Techniques: Real-time Polymerase Chain Reaction

    SiDcR3 sensitizes hepatocellular carcinoma cells through Bcl-2 family members. Notes: ( A ) The prosurvival Bcl-2 family members Bcl-2, Bcl-xL, IAP-2, survivin, and Mcl-1 and proapoptosis member Bax were examined using Western blot analysis after treatment with TRAIL (100 ng/mL) for 24 h. GAPDH was used as loading control. ( B – G ) The ratio of each protein to GAPHD. The differences among different proteins were significant ( P

    Journal: OncoTargets and therapy

    Article Title: Downregulation of DcR3 sensitizes hepatocellular carcinoma cells to TRAIL-induced apoptosis

    doi: 10.2147/OTT.S127202

    Figure Lengend Snippet: SiDcR3 sensitizes hepatocellular carcinoma cells through Bcl-2 family members. Notes: ( A ) The prosurvival Bcl-2 family members Bcl-2, Bcl-xL, IAP-2, survivin, and Mcl-1 and proapoptosis member Bax were examined using Western blot analysis after treatment with TRAIL (100 ng/mL) for 24 h. GAPDH was used as loading control. ( B – G ) The ratio of each protein to GAPHD. The differences among different proteins were significant ( P

    Article Snippet: Rat monoclonal antibodies against Mcl-1, Bcl-XL, Bax, survivin, IAP-2, DR4, DR5, and GAPDH were purchased from Abcam (Shanghai, People’s Republic of China).

    Techniques: Western Blot