anti caspase 9  (Cell Signaling Technology Inc)

 
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  • 99
    Name:
    Caspase 9 Antibody Rat Specific
    Description:
    Caspase 9 ICE LAP6 Mch6 is an important member of the cysteine aspartic acid protease caspase family 1 2 Upon apoptotic stimulation cytochrome c released from mitochondria associates with the 47 kDa procaspase 9 Apaf 1 Apaf 1 mediated activation of caspase 9 involves intrinsic proteolytic processing resulting in cleavage at Asp315 and producing a p35 subunit Another cleavage occurs at Asp330 producing a p37 subunit that can serve to amplify the apoptotic response 3 6 Cleaved caspase 9 further processes other caspase members including caspase 3 and caspase 7 to initiate a caspase cascade which leads to apoptosis 7 10
    Catalog Number:
    9506
    Price:
    None
    Category:
    Primary Antibodies
    Source:
    Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding aspartic acid 353 of rat caspase-9. Antibodies are purified by protein A and peptide affinity chromatography.
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    Structured Review

    Cell Signaling Technology Inc anti caspase 9
    Effect of IP6 on (A) PI3K protein expression, (B) Akt protein expression, (C) pAkt protein expression and (D) <t>caspase-9</t> protein expression in HT-29 cells treated with various concentrations of IP6 (0, 100, 200, 400 μg/mL) for 48 h. The cells were
    Caspase 9 ICE LAP6 Mch6 is an important member of the cysteine aspartic acid protease caspase family 1 2 Upon apoptotic stimulation cytochrome c released from mitochondria associates with the 47 kDa procaspase 9 Apaf 1 Apaf 1 mediated activation of caspase 9 involves intrinsic proteolytic processing resulting in cleavage at Asp315 and producing a p35 subunit Another cleavage occurs at Asp330 producing a p37 subunit that can serve to amplify the apoptotic response 3 6 Cleaved caspase 9 further processes other caspase members including caspase 3 and caspase 7 to initiate a caspase cascade which leads to apoptosis 7 10
    https://www.bioz.com/result/anti caspase 9/product/Cell Signaling Technology Inc
    Average 99 stars, based on 197 article reviews
    Price from $9.99 to $1999.99
    anti caspase 9 - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Inositol hexaphosphate suppresses growth and induces apoptosis in HT-29 colorectal cancer cells in culture: PI3K/Akt pathway as a potential target"

    Article Title: Inositol hexaphosphate suppresses growth and induces apoptosis in HT-29 colorectal cancer cells in culture: PI3K/Akt pathway as a potential target

    Journal: International Journal of Clinical and Experimental Pathology

    doi:

    Effect of IP6 on (A) PI3K protein expression, (B) Akt protein expression, (C) pAkt protein expression and (D) caspase-9 protein expression in HT-29 cells treated with various concentrations of IP6 (0, 100, 200, 400 μg/mL) for 48 h. The cells were
    Figure Legend Snippet: Effect of IP6 on (A) PI3K protein expression, (B) Akt protein expression, (C) pAkt protein expression and (D) caspase-9 protein expression in HT-29 cells treated with various concentrations of IP6 (0, 100, 200, 400 μg/mL) for 48 h. The cells were

    Techniques Used: Expressing

    IP6 decreased the mRNA expression of PI3K and Akt, and it increased the expression of caspase-9 in HT-29 cells
    Figure Legend Snippet: IP6 decreased the mRNA expression of PI3K and Akt, and it increased the expression of caspase-9 in HT-29 cells

    Techniques Used: Expressing

    IP6 decreased the mRNA expression of PI3K and Akt, and it increased the expression of caspase-9 in HT-29 cells
    Figure Legend Snippet: IP6 decreased the mRNA expression of PI3K and Akt, and it increased the expression of caspase-9 in HT-29 cells

    Techniques Used: Expressing

    IP6 decreased the mRNA expression of PI3K and Akt, and it increased the expression of caspase-9 in HT-29 cells
    Figure Legend Snippet: IP6 decreased the mRNA expression of PI3K and Akt, and it increased the expression of caspase-9 in HT-29 cells

    Techniques Used: Expressing

    2) Product Images from "ROS-Mediated Apoptotic Cell Death of Human Colon Cancer LoVo Cells by Milk δ-Valerobetaine"

    Article Title: ROS-Mediated Apoptotic Cell Death of Human Colon Cancer LoVo Cells by Milk δ-Valerobetaine

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-65865-6

    Apoptotic mechanism. ( a,b ) Percentage of apoptosis and representative dot plots of annexin V-FITC and PI-stained cells analyzed by flow cytometry. Data are expressed as mean ± SD of n = 4 experiments. At least 10.000 events were acquired. ( c – h ) Protein expression levels of caspase 3, caspase 9, PARP, Bax and Bcl-2 from LoVo cells treated for 72 h with milk (40% v/v), δVB (2 mM), milk+δVB, or HBSS-10 mM Hepes (40% v/v) (Ctr). Lane 1 = Ctr, lane 2 = milk, lane 3 = δVB, lane 4 = milk + δVB. Analysis of densitometric intensity was calculated with Image J 1.52n version software. Arbitrary units of protein expression (AU) were quantified using α-tubulin or β-actin. Antibodies against Bax, Bcl-2 and SIRT6 (reported in Fig. 4 ) were blotted on the same filter and quantified by using the same loading control (α-tubulin). ( i,j ) Flow cytometric analysis and representative dot plots of annexin V-FITC and PI double staining LoVo cells treated with caspase 9 inhibitor Z-LEHD-FMK (40 μM) or chloroquine (50 μM). Data are expressed as mean ± SD of n = 3 experiments. At least 10.000 events were acquired. ( k ) Cleaved caspase 3 protein expression level from LoVo cells treated for 72 h with milk+δVB, Z-LEHD-FMK + milk+δVB or HBSS-10 mM Hepes (40% v/v) (Ctr). Lane 1 = Ctr, lane 2 = milk + δVB, lane 3 = Z-LEHD-FMK + milk+δVB. ( l ) Protein expression levels of caspase 8 in LoVo cells after 72 h of treatment with milk (40% v/v), δVB (2 mM), milk+δVB, or HBSS-10 mM Hepes (40% v/v) (Ctr). Lane 1=Ctr, lane 2 = milk, lane 3 = δVB, lane 4 = milk + δVB. * P
    Figure Legend Snippet: Apoptotic mechanism. ( a,b ) Percentage of apoptosis and representative dot plots of annexin V-FITC and PI-stained cells analyzed by flow cytometry. Data are expressed as mean ± SD of n = 4 experiments. At least 10.000 events were acquired. ( c – h ) Protein expression levels of caspase 3, caspase 9, PARP, Bax and Bcl-2 from LoVo cells treated for 72 h with milk (40% v/v), δVB (2 mM), milk+δVB, or HBSS-10 mM Hepes (40% v/v) (Ctr). Lane 1 = Ctr, lane 2 = milk, lane 3 = δVB, lane 4 = milk + δVB. Analysis of densitometric intensity was calculated with Image J 1.52n version software. Arbitrary units of protein expression (AU) were quantified using α-tubulin or β-actin. Antibodies against Bax, Bcl-2 and SIRT6 (reported in Fig. 4 ) were blotted on the same filter and quantified by using the same loading control (α-tubulin). ( i,j ) Flow cytometric analysis and representative dot plots of annexin V-FITC and PI double staining LoVo cells treated with caspase 9 inhibitor Z-LEHD-FMK (40 μM) or chloroquine (50 μM). Data are expressed as mean ± SD of n = 3 experiments. At least 10.000 events were acquired. ( k ) Cleaved caspase 3 protein expression level from LoVo cells treated for 72 h with milk+δVB, Z-LEHD-FMK + milk+δVB or HBSS-10 mM Hepes (40% v/v) (Ctr). Lane 1 = Ctr, lane 2 = milk + δVB, lane 3 = Z-LEHD-FMK + milk+δVB. ( l ) Protein expression levels of caspase 8 in LoVo cells after 72 h of treatment with milk (40% v/v), δVB (2 mM), milk+δVB, or HBSS-10 mM Hepes (40% v/v) (Ctr). Lane 1=Ctr, lane 2 = milk, lane 3 = δVB, lane 4 = milk + δVB. * P

    Techniques Used: Staining, Flow Cytometry, Expressing, Software, Double Staining

    3) Product Images from "Tumor- and mitochondria-targeted nanoparticles eradicate drug resistant lung cancer through mitochondrial pathway of apoptosis"

    Article Title: Tumor- and mitochondria-targeted nanoparticles eradicate drug resistant lung cancer through mitochondrial pathway of apoptosis

    Journal: Journal of Nanobiotechnology

    doi: 10.1186/s12951-019-0562-3

    a Cell apoptosis rate detected by flow cytometry. A549 and A549/ADR cells were treated with different formulations that contained a total PTX concentration of 10 μM for 24 h. b Proteins involved in the apoptosis signaling pathways in A549 and A549/ADR cells as determined by Western blotting. ① Control (PBS); ② Taxol; ③ TP/PTX; and ④ TPH/PTX nanomicells. Activity ratios of caspase-3 and caspase-9 and expression ratios of the pro-apoptotic proteins Bax and the anti-apoptotic proteins Bcl-2 in A549 and A549/ADR cells after incubation with the various formulations. β-actin was also assessed by Western blotting. All protein levels were quantified densitometrically and normalized to β-actin. All data are presented as the means ± standard deviations (n = 3); (1) Image of western blot; (2) Grey level of western blot. * P
    Figure Legend Snippet: a Cell apoptosis rate detected by flow cytometry. A549 and A549/ADR cells were treated with different formulations that contained a total PTX concentration of 10 μM for 24 h. b Proteins involved in the apoptosis signaling pathways in A549 and A549/ADR cells as determined by Western blotting. ① Control (PBS); ② Taxol; ③ TP/PTX; and ④ TPH/PTX nanomicells. Activity ratios of caspase-3 and caspase-9 and expression ratios of the pro-apoptotic proteins Bax and the anti-apoptotic proteins Bcl-2 in A549 and A549/ADR cells after incubation with the various formulations. β-actin was also assessed by Western blotting. All protein levels were quantified densitometrically and normalized to β-actin. All data are presented as the means ± standard deviations (n = 3); (1) Image of western blot; (2) Grey level of western blot. * P

    Techniques Used: Flow Cytometry, Cytometry, Concentration Assay, Western Blot, Activity Assay, Expressing, Incubation

    4) Product Images from "Tom70 Mediates Sendai Virus-Induced Apoptosis on Mitochondria"

    Article Title: Tom70 Mediates Sendai Virus-Induced Apoptosis on Mitochondria

    Journal: Journal of Virology

    doi: 10.1128/JVI.02959-14

    Sendai virus-induced apoptosis depends on caspase-9. (A) HEK293 cells were infected with Sendai virus at an MOI of 0.4, 0.8, or 1.2 for 24 h (left) or at an MOI of 1 for 18, 21, or 24 h (right). Cell death was quantified using annexin V/PI double staining
    Figure Legend Snippet: Sendai virus-induced apoptosis depends on caspase-9. (A) HEK293 cells were infected with Sendai virus at an MOI of 0.4, 0.8, or 1.2 for 24 h (left) or at an MOI of 1 for 18, 21, or 24 h (right). Cell death was quantified using annexin V/PI double staining

    Techniques Used: Infection, Double Staining

    5) Product Images from "Hypophosphatemia leads to rickets by impairing caspase-mediated apoptosis of hypertrophic chondrocytes"

    Article Title: Hypophosphatemia leads to rickets by impairing caspase-mediated apoptosis of hypertrophic chondrocytes

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.0502249102

    In vivo inhibition of caspase-3 or caspase-9 leads to expansion of the hypertrophic chondrocyte layer. Shown are hematoxylin/eosin-stained sections from 24-day-old mice injected at days 18–23 with DMSO, caspase-3 inhibitor (Z-DEVD-FMK), or caspase-9
    Figure Legend Snippet: In vivo inhibition of caspase-3 or caspase-9 leads to expansion of the hypertrophic chondrocyte layer. Shown are hematoxylin/eosin-stained sections from 24-day-old mice injected at days 18–23 with DMSO, caspase-3 inhibitor (Z-DEVD-FMK), or caspase-9

    Techniques Used: In Vivo, Inhibition, Staining, Mouse Assay, Injection

    6) Product Images from "Caspase-3 Inhibition Attenuates the Cytopathic Effects of EV71 Infection"

    Article Title: Caspase-3 Inhibition Attenuates the Cytopathic Effects of EV71 Infection

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.00817

    Non-structural 3C protease binds caspase-8 and caspase-9. (A) Cell morphologic analysis and nuclear morphologic analysis were performed in HepG2 cells because they are easy to transfect. Analyses were performed 36 h after transfection with 4 μg of pEGFP-3C (3C) or the corresponding control vector pEGFP (Vector). Bar = 20 μm. (B) The expression and activity of caspase-3, caspase-8, and caspase-9 proteins after transfection of HepG2 cells with VR1012-3C-HA or the corresponding control vector VR1012-HA (Vec) was assessed at 36 h by Western blot analysis (top) and by activity assay (lower). Histone is shown as a loading control. Luminescence values in each cell were calculated and normalized to 1.0 in vector-transfected cells. The results indicate the means ± SD of three independent experiments. ∗∗∗ P
    Figure Legend Snippet: Non-structural 3C protease binds caspase-8 and caspase-9. (A) Cell morphologic analysis and nuclear morphologic analysis were performed in HepG2 cells because they are easy to transfect. Analyses were performed 36 h after transfection with 4 μg of pEGFP-3C (3C) or the corresponding control vector pEGFP (Vector). Bar = 20 μm. (B) The expression and activity of caspase-3, caspase-8, and caspase-9 proteins after transfection of HepG2 cells with VR1012-3C-HA or the corresponding control vector VR1012-HA (Vec) was assessed at 36 h by Western blot analysis (top) and by activity assay (lower). Histone is shown as a loading control. Luminescence values in each cell were calculated and normalized to 1.0 in vector-transfected cells. The results indicate the means ± SD of three independent experiments. ∗∗∗ P

    Techniques Used: Transfection, Plasmid Preparation, Expressing, Activity Assay, Western Blot

    7) Product Images from "Decay accelerating factor (CD55) protects neuronal cells from chemical hypoxia-induced injury"

    Article Title: Decay accelerating factor (CD55) protects neuronal cells from chemical hypoxia-induced injury

    Journal: Journal of Neuroinflammation

    doi: 10.1186/1742-2094-7-24

    DAF suppresses the expression of activated caspase in neurons exposed to chemical hypoxia . Primary rat cortical neurons were exposed to glucose free medium containing 1.5 mM NaCN for 1 hr then allowed to recover overnight in normal medium with or without DAF. (a) Cell lysates were analyzed by immunoblot using anti-active caspase-3, anti-caspase-9, and anti-β-actin antibodies. (b) Data was quantified by densitometry (n = 10). (c and d) Cells were marked with anti-active caspase-3 (red), anti-MAC (green) antibodies, and DAPI (blue) (n = 3). Scale bar = 50 μm. (one-way ANOVA followed by Newman-Keuls test. * p
    Figure Legend Snippet: DAF suppresses the expression of activated caspase in neurons exposed to chemical hypoxia . Primary rat cortical neurons were exposed to glucose free medium containing 1.5 mM NaCN for 1 hr then allowed to recover overnight in normal medium with or without DAF. (a) Cell lysates were analyzed by immunoblot using anti-active caspase-3, anti-caspase-9, and anti-β-actin antibodies. (b) Data was quantified by densitometry (n = 10). (c and d) Cells were marked with anti-active caspase-3 (red), anti-MAC (green) antibodies, and DAPI (blue) (n = 3). Scale bar = 50 μm. (one-way ANOVA followed by Newman-Keuls test. * p

    Techniques Used: Expressing

    8) Product Images from "Adenoviruses-mediated transduction of human oesophageal carcinoma cells with the interferon-λ genes produced anti-tumour effects"

    Article Title: Adenoviruses-mediated transduction of human oesophageal carcinoma cells with the interferon-λ genes produced anti-tumour effects

    Journal: British Journal of Cancer

    doi: 10.1038/bjc.2011.379

    Expressions of apoptosis-linked proteins with Ad/IFN- λ treatments. YES-2 and T.Tn cells were infected with Ad (1000 MOI) and cultured for 72 h. Expressions of caspase-3, PARP, the respective cleaved forms ( A ), caspase-8, caspase-9, the respective cleaved forms, Bax, Bcl-2 and cytochrome C in a cytoplasmic fraction ( B ) were analysed with western blot analyses. Actin and GAPDH expression are shown as a loading control for total, cytoplasmic protein, respectively.
    Figure Legend Snippet: Expressions of apoptosis-linked proteins with Ad/IFN- λ treatments. YES-2 and T.Tn cells were infected with Ad (1000 MOI) and cultured for 72 h. Expressions of caspase-3, PARP, the respective cleaved forms ( A ), caspase-8, caspase-9, the respective cleaved forms, Bax, Bcl-2 and cytochrome C in a cytoplasmic fraction ( B ) were analysed with western blot analyses. Actin and GAPDH expression are shown as a loading control for total, cytoplasmic protein, respectively.

    Techniques Used: Infection, Cell Culture, Western Blot, Expressing

    9) Product Images from "Irisin Promotes Human Umbilical Vein Endothelial Cell Proliferation through the ERK Signaling Pathway and Partly Suppresses High Glucose-Induced Apoptosis"

    Article Title: Irisin Promotes Human Umbilical Vein Endothelial Cell Proliferation through the ERK Signaling Pathway and Partly Suppresses High Glucose-Induced Apoptosis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0110273

    Irisin mediates Bax,Bcl-2,GSK-3β, Caspase-9 and Caspase-3 protein levels in HUVECs. HUVECs were cultured with or without irisin (20 nM) for 24 h. Bax, Bcl-2, Bad, GSK-3β, Caspase-9 and Caspase-3 in cell lysates were analyzed by Western blot. (B) Densitometric analysis of the related bands was expressed as the relative optical band density and was corrected using respective total proteins as loading controls and normalized against the untreated control. The data were expressed as the mean ± SE of three independent experiments, *p
    Figure Legend Snippet: Irisin mediates Bax,Bcl-2,GSK-3β, Caspase-9 and Caspase-3 protein levels in HUVECs. HUVECs were cultured with or without irisin (20 nM) for 24 h. Bax, Bcl-2, Bad, GSK-3β, Caspase-9 and Caspase-3 in cell lysates were analyzed by Western blot. (B) Densitometric analysis of the related bands was expressed as the relative optical band density and was corrected using respective total proteins as loading controls and normalized against the untreated control. The data were expressed as the mean ± SE of three independent experiments, *p

    Techniques Used: Cell Culture, Western Blot

    10) Product Images from "Mitophagy switches cell death from apoptosis to necrosis in NSCLC cells treated with oncolytic measles virus"

    Article Title: Mitophagy switches cell death from apoptosis to necrosis in NSCLC cells treated with oncolytic measles virus

    Journal: Oncotarget

    doi:

    Autophagy protects NSCLCs from apoptosis leading to enhanced viral replication (a) A549 and H1299 cells were infected with MV-Edm at a MOI of 0.5 or 2 in the absence or presence of a pan-caspase inhibitor z-VAD-fmk (80μM) for 72 h. Cell viability was quantified by trypan-blue exclusion. Means + SD of triplicates are shown. Similar results were obtained in three independent experiments. (b) A549 and H1299 cells were infected with MV-Edm at a MOI of 0.5 for 24, 48 and 72 h. Cell lysates were harvested for Western blot. Cell lysates from untreated and staurosporin (STS) treated cells were used as negative and positive controls, respectively. Representative blots from two independent experiments are shown. (c) Cytoplasmic cytochrome, cleaved caspase -9, -3 and PARP were evaluated by Western blotting cell lysates harvested from A549 and H1299 cells transfected with siRNAs targeting ATG7, BECN1 or non-specific control siRNA followed by MV-Edm infection at a MOI of 0.5 for 48 h. A representative result from two independent experiments is shown. (d) A549 cells were transfected with siRNA targeting ATG7, BECN1, SQSTM1, or with non-targeting control siRNA for 24 h followed by infection with MV-Edm (MOI = 0.5) for another 48 h. Cells were stained by DAPI before subjected to fluorescent confocal microscopy for evaluation of fragmented nuclei (blue spots). Cells treated with staurosporin (STS, 500 nM) for 12 h were used as a positive control. Scale bars represent 40 μm (upper two panels) and 10 μm (lower panel). Dashed white lines highlight multinucleated syncytia. (e) A549 cells were transfected with siRNA targeting ATG7, BECN1, SQSTM1, or with non-targeting control siRNA for 24 h followed by infection with MV-Edm (MOI = 0.5) in the presence or absence of zVAD (80 μM) for another 48 h. Cells were harvested and the specific apoptosis was analyzed by determining hypodiploid nuclei by FACS. Means + SD of triplicates are shown. Similar results were obtained in three independent experiments. (f) A549 cells were transfected with siRNAs targeting ATG7, BECN1, SQSTM1 or non-specific control siRNA for 24 h followed by infection with MV-Edm (MOI = 0.5) in the absence or presence of zVAD (80 μM) for another 48 h. Then the expression of N- and H-viral structural genes was quantified by qRT-PCR. Means + SD of triplicates are shown. Similar results were obtained in three independent experiments. # p  >  0.05, * p
    Figure Legend Snippet: Autophagy protects NSCLCs from apoptosis leading to enhanced viral replication (a) A549 and H1299 cells were infected with MV-Edm at a MOI of 0.5 or 2 in the absence or presence of a pan-caspase inhibitor z-VAD-fmk (80μM) for 72 h. Cell viability was quantified by trypan-blue exclusion. Means + SD of triplicates are shown. Similar results were obtained in three independent experiments. (b) A549 and H1299 cells were infected with MV-Edm at a MOI of 0.5 for 24, 48 and 72 h. Cell lysates were harvested for Western blot. Cell lysates from untreated and staurosporin (STS) treated cells were used as negative and positive controls, respectively. Representative blots from two independent experiments are shown. (c) Cytoplasmic cytochrome, cleaved caspase -9, -3 and PARP were evaluated by Western blotting cell lysates harvested from A549 and H1299 cells transfected with siRNAs targeting ATG7, BECN1 or non-specific control siRNA followed by MV-Edm infection at a MOI of 0.5 for 48 h. A representative result from two independent experiments is shown. (d) A549 cells were transfected with siRNA targeting ATG7, BECN1, SQSTM1, or with non-targeting control siRNA for 24 h followed by infection with MV-Edm (MOI = 0.5) for another 48 h. Cells were stained by DAPI before subjected to fluorescent confocal microscopy for evaluation of fragmented nuclei (blue spots). Cells treated with staurosporin (STS, 500 nM) for 12 h were used as a positive control. Scale bars represent 40 μm (upper two panels) and 10 μm (lower panel). Dashed white lines highlight multinucleated syncytia. (e) A549 cells were transfected with siRNA targeting ATG7, BECN1, SQSTM1, or with non-targeting control siRNA for 24 h followed by infection with MV-Edm (MOI = 0.5) in the presence or absence of zVAD (80 μM) for another 48 h. Cells were harvested and the specific apoptosis was analyzed by determining hypodiploid nuclei by FACS. Means + SD of triplicates are shown. Similar results were obtained in three independent experiments. (f) A549 cells were transfected with siRNAs targeting ATG7, BECN1, SQSTM1 or non-specific control siRNA for 24 h followed by infection with MV-Edm (MOI = 0.5) in the absence or presence of zVAD (80 μM) for another 48 h. Then the expression of N- and H-viral structural genes was quantified by qRT-PCR. Means + SD of triplicates are shown. Similar results were obtained in three independent experiments. # p > 0.05, * p

    Techniques Used: Infection, Western Blot, Transfection, Staining, Confocal Microscopy, Positive Control, FACS, Expressing, Quantitative RT-PCR

    11) Product Images from "Overexpression of Forkhead Box Protein M1 (FOXM1) in Ovarian Cancer Correlates with Poor Patient Survival and Contributes to Paclitaxel Resistance"

    Article Title: Overexpression of Forkhead Box Protein M1 (FOXM1) in Ovarian Cancer Correlates with Poor Patient Survival and Contributes to Paclitaxel Resistance

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0113478

    Paclitaxel treatment down-regulates FOXM1 expression in SKOV-3 but not in SKOV3-TR cells. The paclitaxel sensitive SKOV-3 and resistant SKOV3-TR ovarian cancer cells were treated with 100 nM paclitaxel and harvested at times indicated for Western blot analysis. Paclitaxel treatment down-regulated FOXM1 expression at time points 48 h and 72 h in SKOV-3 but not in SKOV3-TR as shown by immunoblotting. There were also no marked changes in the cleaved Caspase-9 and Caspase-7 expression.
    Figure Legend Snippet: Paclitaxel treatment down-regulates FOXM1 expression in SKOV-3 but not in SKOV3-TR cells. The paclitaxel sensitive SKOV-3 and resistant SKOV3-TR ovarian cancer cells were treated with 100 nM paclitaxel and harvested at times indicated for Western blot analysis. Paclitaxel treatment down-regulated FOXM1 expression at time points 48 h and 72 h in SKOV-3 but not in SKOV3-TR as shown by immunoblotting. There were also no marked changes in the cleaved Caspase-9 and Caspase-7 expression.

    Techniques Used: Expressing, Western Blot

    12) Product Images from "A delay prior to mitotic entry triggers caspase 8-dependent cell death in p53-deficient Hela and HCT-116 cells"

    Article Title: A delay prior to mitotic entry triggers caspase 8-dependent cell death in p53-deficient Hela and HCT-116 cells

    Journal: Cell Cycle

    doi: 10.1080/15384101.2015.1007781

    Initiator caspase 9 is not required for cell death in stathmin-deleted Hela cells. ( A ) Western blot demonstrating knockdown of caspase 9 alone or in combination with stathmin depletion. Tubulin was used as a loading control. ( B ) Percent trypan blue positive cells for the indicated conditions measured 48 hr after transfection. * denotes P
    Figure Legend Snippet: Initiator caspase 9 is not required for cell death in stathmin-deleted Hela cells. ( A ) Western blot demonstrating knockdown of caspase 9 alone or in combination with stathmin depletion. Tubulin was used as a loading control. ( B ) Percent trypan blue positive cells for the indicated conditions measured 48 hr after transfection. * denotes P

    Techniques Used: Western Blot, Transfection

    13) Product Images from "Antitumor and apoptosis-inducing effects of α-mangostin extracted from the pericarp of the mangosteen fruit (Garcinia mangostana L.) in YD-15 tongue mucoepidermoid carcinoma cells"

    Article Title: Antitumor and apoptosis-inducing effects of α-mangostin extracted from the pericarp of the mangosteen fruit (Garcinia mangostana L.) in YD-15 tongue mucoepidermoid carcinoma cells

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2016.2517

    Effects of α-mangostin on apoptosis. (A) YD-15 cells were treated with α-mangostin (0, 10 and 15 µ M) for 24 h. Cell lysates were prepared as described in the Material and methods and analyzed by 12% SDS-PAGE, followed by western blot analysis. Membranes were incubated with anti-caspase-3, anti-caspase-9 and anti-PARP antibodies. Blots were also probed with anti-β-actin antibody to confirm equal sample loading. (B) Nude mice were treated with α-mangostin (0, 10 and 20 mg/kg) for 22 days. To identify cleaved (c)-caspase-3 protein, immunohistochemcal staining was performed on tumor tissues collected from mouse xenografts as described in the Material and methods. Cleaved caspase-3 was observed under a microscope and photographed (×200 magnification). Paraffin-embedded tumors were sectioned to 5- µ m thickness.
    Figure Legend Snippet: Effects of α-mangostin on apoptosis. (A) YD-15 cells were treated with α-mangostin (0, 10 and 15 µ M) for 24 h. Cell lysates were prepared as described in the Material and methods and analyzed by 12% SDS-PAGE, followed by western blot analysis. Membranes were incubated with anti-caspase-3, anti-caspase-9 and anti-PARP antibodies. Blots were also probed with anti-β-actin antibody to confirm equal sample loading. (B) Nude mice were treated with α-mangostin (0, 10 and 20 mg/kg) for 22 days. To identify cleaved (c)-caspase-3 protein, immunohistochemcal staining was performed on tumor tissues collected from mouse xenografts as described in the Material and methods. Cleaved caspase-3 was observed under a microscope and photographed (×200 magnification). Paraffin-embedded tumors were sectioned to 5- µ m thickness.

    Techniques Used: SDS Page, Western Blot, Incubation, Mouse Assay, Staining, Microscopy

    14) Product Images from "Combined expression of miR-34a and Smac mediated by oncolytic vaccinia virus synergistically promote anti-tumor effects in Multiple Myeloma"

    Article Title: Combined expression of miR-34a and Smac mediated by oncolytic vaccinia virus synergistically promote anti-tumor effects in Multiple Myeloma

    Journal: Scientific Reports

    doi: 10.1038/srep32174

    The mechanism schematic mode of viral oncolysis mediated by VV-miR-34a and VV-Smac is shown. ( A ) OVVs infect the tumor cells and elicit the tumor specific oncolytic effect. ( B ) miR-34a/Smac mediated caspase-9-dependent apoptosis pathway. Enforced expression of miR-34a by OVV blocks the function of Bcl-2, which promotes the release of cytochrome c from mitochondrial endomembrane. On the other hand, overexpression of Smac by OVV inhibits the function of inhibitors of apoptosis proteins (c-IAP, XIAP). Both of them synergistically activate the caspase-9 induced apoptosis pathway. This signal pathway image was drawn by Wen Lei.
    Figure Legend Snippet: The mechanism schematic mode of viral oncolysis mediated by VV-miR-34a and VV-Smac is shown. ( A ) OVVs infect the tumor cells and elicit the tumor specific oncolytic effect. ( B ) miR-34a/Smac mediated caspase-9-dependent apoptosis pathway. Enforced expression of miR-34a by OVV blocks the function of Bcl-2, which promotes the release of cytochrome c from mitochondrial endomembrane. On the other hand, overexpression of Smac by OVV inhibits the function of inhibitors of apoptosis proteins (c-IAP, XIAP). Both of them synergistically activate the caspase-9 induced apoptosis pathway. This signal pathway image was drawn by Wen Lei.

    Techniques Used: Expressing, Over Expression

    15) Product Images from "The effect of aloe-emodin-induced photodynamic activity on the apoptosis of human gastric cancer cells: A pilot study"

    Article Title: The effect of aloe-emodin-induced photodynamic activity on the apoptosis of human gastric cancer cells: A pilot study

    Journal: Oncology Letters

    doi: 10.3892/ol.2017.5915

    Protein levels of caspase-3 and caspase-9 in group I (without AE and illumination), group II (10 µM AE), group III (illumination, 12.8 J/cm 2 ) and group IV (photodynamic therapy), as determined by western blot. AE, aloe-emodin.
    Figure Legend Snippet: Protein levels of caspase-3 and caspase-9 in group I (without AE and illumination), group II (10 µM AE), group III (illumination, 12.8 J/cm 2 ) and group IV (photodynamic therapy), as determined by western blot. AE, aloe-emodin.

    Techniques Used: Western Blot

    16) Product Images from "Mangiferin attenuates oxidative stress induced renal cell damage through activation of PI3K induced Akt and Nrf-2 mediated signaling pathways"

    Article Title: Mangiferin attenuates oxidative stress induced renal cell damage through activation of PI3K induced Akt and Nrf-2 mediated signaling pathways

    Journal: Biochemistry and Biophysics Reports

    doi: 10.1016/j.bbrep.2016.01.011

    (A) Immunoblot analysis of Bax and Bcl2 in response to tBHP and mangiferin exposure, in NKE cells. (B) Immunoblot analysis of Caspase 8, tBid, Bax, cytochrome C, Apaf-1, Caspase 9, Caspase 3 and PARP. NKE cells were exposed to tBHP (50 µM) and mangiferin (20 µM) according to the defined protocol. β actin was used as an internal control Control: untreated cells, M: exposed to mangiferin, tBHP: exposed to tBHP for 18 h, M+tBHP: Mangiferin treated for 2 h followed by 18 h of tBHP exposure. For inhibitor studies, M+T+I: cells were treated with LY294002(I) follwed by Mangiferin(M) and Tbhp(T). All data are mean±SD, of 3 independent experiments and were analyzed by one-way ANOVA. “*” represents the significant difference between the normal control and tBHP exposed cells and “#” represents the significant difference between the tBHP exposed and tBHP exposed antioxidant treated cells ( P *
    Figure Legend Snippet: (A) Immunoblot analysis of Bax and Bcl2 in response to tBHP and mangiferin exposure, in NKE cells. (B) Immunoblot analysis of Caspase 8, tBid, Bax, cytochrome C, Apaf-1, Caspase 9, Caspase 3 and PARP. NKE cells were exposed to tBHP (50 µM) and mangiferin (20 µM) according to the defined protocol. β actin was used as an internal control Control: untreated cells, M: exposed to mangiferin, tBHP: exposed to tBHP for 18 h, M+tBHP: Mangiferin treated for 2 h followed by 18 h of tBHP exposure. For inhibitor studies, M+T+I: cells were treated with LY294002(I) follwed by Mangiferin(M) and Tbhp(T). All data are mean±SD, of 3 independent experiments and were analyzed by one-way ANOVA. “*” represents the significant difference between the normal control and tBHP exposed cells and “#” represents the significant difference between the tBHP exposed and tBHP exposed antioxidant treated cells ( P *

    Techniques Used:

    17) Product Images from "Inhibitory effect of hyperoside isolated from Zanthoxylum bungeanum leaves on SW620 human colorectal cancer cells via induction of the p53 signaling pathway and apoptosis"

    Article Title: Inhibitory effect of hyperoside isolated from Zanthoxylum bungeanum leaves on SW620 human colorectal cancer cells via induction of the p53 signaling pathway and apoptosis

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2017.6710

    Effect of Zanthoxylum bungeanum on p53-associated proteins. SW620 cells were exposed to 12.5, 25 and 50 µM hyperoside for 48 h. (A) Protein expression of p53-associated proteins. Quantification of (B) p53, (C) p21, (D) Bax, (E) Cytc, (F) caspase-9, (G) caspase-3 and (H) Apaf-1. *P
    Figure Legend Snippet: Effect of Zanthoxylum bungeanum on p53-associated proteins. SW620 cells were exposed to 12.5, 25 and 50 µM hyperoside for 48 h. (A) Protein expression of p53-associated proteins. Quantification of (B) p53, (C) p21, (D) Bax, (E) Cytc, (F) caspase-9, (G) caspase-3 and (H) Apaf-1. *P

    Techniques Used: Expressing

    18) Product Images from "Caspase-3 Inhibition Attenuates the Cytopathic Effects of EV71 Infection"

    Article Title: Caspase-3 Inhibition Attenuates the Cytopathic Effects of EV71 Infection

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.00817

    Non-structural 3C protease binds caspase-8 and caspase-9. (A) Cell morphologic analysis and nuclear morphologic analysis were performed in HepG2 cells because they are easy to transfect. Analyses were performed 36 h after transfection with 4 μg of pEGFP-3C (3C) or the corresponding control vector pEGFP (Vector). Bar = 20 μm. (B) The expression and activity of caspase-3, caspase-8, and caspase-9 proteins after transfection of HepG2 cells with VR1012-3C-HA or the corresponding control vector VR1012-HA (Vec) was assessed at 36 h by Western blot analysis (top) and by activity assay (lower). Histone is shown as a loading control. Luminescence values in each cell were calculated and normalized to 1.0 in vector-transfected cells. The results indicate the means ± SD of three independent experiments. ∗∗∗ P
    Figure Legend Snippet: Non-structural 3C protease binds caspase-8 and caspase-9. (A) Cell morphologic analysis and nuclear morphologic analysis were performed in HepG2 cells because they are easy to transfect. Analyses were performed 36 h after transfection with 4 μg of pEGFP-3C (3C) or the corresponding control vector pEGFP (Vector). Bar = 20 μm. (B) The expression and activity of caspase-3, caspase-8, and caspase-9 proteins after transfection of HepG2 cells with VR1012-3C-HA or the corresponding control vector VR1012-HA (Vec) was assessed at 36 h by Western blot analysis (top) and by activity assay (lower). Histone is shown as a loading control. Luminescence values in each cell were calculated and normalized to 1.0 in vector-transfected cells. The results indicate the means ± SD of three independent experiments. ∗∗∗ P

    Techniques Used: Transfection, Plasmid Preparation, Expressing, Activity Assay, Western Blot

    19) Product Images from "Long non-coding RNA C5orf66-AS1 prevents oral squamous cell carcinoma through inhibiting cell growth and metastasis"

    Article Title: Long non-coding RNA C5orf66-AS1 prevents oral squamous cell carcinoma through inhibiting cell growth and metastasis

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3913

    Effect of lncRNA C5orf66-AS1-plasmid on Bcl-2, Bax, MMP9, Caspase-3, Caspase-7 and Caspase-9 expression levels in SCC9 cells. (A) Western blot analysis was performed at 48 h after cell transfection to detect the protein levels. (B) Bcl-2, (C) Bax, (D) MMP9, (E) cleaved Caspase-3/Caspase-3, (F) cleaved Caspase-7/Caspase-7 and (G) cleaved Caspase-9/Caspase-9 in SCC9 cells were detected using reverse transcription-quantitative polymerase chain reaction. ** P
    Figure Legend Snippet: Effect of lncRNA C5orf66-AS1-plasmid on Bcl-2, Bax, MMP9, Caspase-3, Caspase-7 and Caspase-9 expression levels in SCC9 cells. (A) Western blot analysis was performed at 48 h after cell transfection to detect the protein levels. (B) Bcl-2, (C) Bax, (D) MMP9, (E) cleaved Caspase-3/Caspase-3, (F) cleaved Caspase-7/Caspase-7 and (G) cleaved Caspase-9/Caspase-9 in SCC9 cells were detected using reverse transcription-quantitative polymerase chain reaction. ** P

    Techniques Used: Plasmid Preparation, Expressing, Western Blot, Transfection, Real-time Polymerase Chain Reaction

    Effect of lncRNA C5orf66-AS1-siRNA on Bcl-2, Bax, MMP9, Caspase-3, Caspase-7 and Caspase-9 expression levels in SCC9 cells. (A) Western blot analysis was performed at 48 h after cell transfection to detect the protein levels. (B) Bcl-2, (C) Bax, (D) MMP9, (E) cleaved Caspase-3/Caspase-3, (F) cleaved Caspase-7/Caspase-7 and (G) cleaved Caspase-9/Caspase-9 in SCC9 cells were detected using transcription-quantitative polymerase chain reaction. ** P
    Figure Legend Snippet: Effect of lncRNA C5orf66-AS1-siRNA on Bcl-2, Bax, MMP9, Caspase-3, Caspase-7 and Caspase-9 expression levels in SCC9 cells. (A) Western blot analysis was performed at 48 h after cell transfection to detect the protein levels. (B) Bcl-2, (C) Bax, (D) MMP9, (E) cleaved Caspase-3/Caspase-3, (F) cleaved Caspase-7/Caspase-7 and (G) cleaved Caspase-9/Caspase-9 in SCC9 cells were detected using transcription-quantitative polymerase chain reaction. ** P

    Techniques Used: Expressing, Western Blot, Transfection, Real-time Polymerase Chain Reaction

    20) Product Images from "Triptolide Induces S Phase Arrest and Apoptosis in Gallbladder Cancer Cells"

    Article Title: Triptolide Induces S Phase Arrest and Apoptosis in Gallbladder Cancer Cells

    Journal: Molecules

    doi: 10.3390/molecules19022612

    Triptolide regulates the expression of apoptosis-related proteins in gallbladder cancer cells. Expression of cleaved-caspase-3, cleaved-caspase-9, cleaved-PARP, Bax, and Bcl-2 in GBC-SD cells ( A ) and SGC-996 cells ( B ) treated with triptolide at the indicated doses for 48h. β-actin was used as a loading control. ( C ) and ( D ), Ratio of Bcl-2 to Bax determined by band density, shown as mean ± SD, compared with the control (designated as 1.00). * p
    Figure Legend Snippet: Triptolide regulates the expression of apoptosis-related proteins in gallbladder cancer cells. Expression of cleaved-caspase-3, cleaved-caspase-9, cleaved-PARP, Bax, and Bcl-2 in GBC-SD cells ( A ) and SGC-996 cells ( B ) treated with triptolide at the indicated doses for 48h. β-actin was used as a loading control. ( C ) and ( D ), Ratio of Bcl-2 to Bax determined by band density, shown as mean ± SD, compared with the control (designated as 1.00). * p

    Techniques Used: Expressing

    21) Product Images from "Bcl-2 family inhibition sensitizes human prostate cancer cells to docetaxel and promotes unexpected apoptosis under caspase-9 inhibition"

    Article Title: Bcl-2 family inhibition sensitizes human prostate cancer cells to docetaxel and promotes unexpected apoptosis under caspase-9 inhibition

    Journal: Oncotarget

    doi:

    Inhibition of caspase-9 promotes apoptosis in ABT-263-treated PC3 cells (A) PC3 cells were treated with both DTX (2.5 nM) and ABT-263 (4 μM). After 24 h, cells were stained with FITC-conjugated Annexin V and PI, and flow cytometry was performed. The numbers represent the percentages of each subset. (B) PC3 cells were treated with DTX (2.5 nM) and/or ABT-263 (4 μM). After 24 h, cells were harvested and cell lysates assayed for their expression of caspase-3, -8, -9, and -2 by immunoblot. β-actin was used as a loading control. (C) PC3 cells were treated with both DTX (2.5 nM) and ABT-263 (4 μM) in the presence of the indicated caspase inhibitors. After 24 h, flow cytometry was performed as described previously. The numbers represent the percentages of each subset. (D) PC3 cells were treated with either DTX (2.5 nM) or ABT-263 (4 μM) in the presence of the indicated caspase inhibitors. DTX, docetaxel; panCi, pan-caspase inhibitor; C9i, caspase-9 inhibitor; C8i, caspase-8 inhibitor; C2i, caspase-2 inhibitor. As the vehicle control, the same volume of DMSO was added.
    Figure Legend Snippet: Inhibition of caspase-9 promotes apoptosis in ABT-263-treated PC3 cells (A) PC3 cells were treated with both DTX (2.5 nM) and ABT-263 (4 μM). After 24 h, cells were stained with FITC-conjugated Annexin V and PI, and flow cytometry was performed. The numbers represent the percentages of each subset. (B) PC3 cells were treated with DTX (2.5 nM) and/or ABT-263 (4 μM). After 24 h, cells were harvested and cell lysates assayed for their expression of caspase-3, -8, -9, and -2 by immunoblot. β-actin was used as a loading control. (C) PC3 cells were treated with both DTX (2.5 nM) and ABT-263 (4 μM) in the presence of the indicated caspase inhibitors. After 24 h, flow cytometry was performed as described previously. The numbers represent the percentages of each subset. (D) PC3 cells were treated with either DTX (2.5 nM) or ABT-263 (4 μM) in the presence of the indicated caspase inhibitors. DTX, docetaxel; panCi, pan-caspase inhibitor; C9i, caspase-9 inhibitor; C8i, caspase-8 inhibitor; C2i, caspase-2 inhibitor. As the vehicle control, the same volume of DMSO was added.

    Techniques Used: Inhibition, Staining, Flow Cytometry, Cytometry, Expressing

    Induction of caspase-8-dependent ABT-263-induced apoptosis of LNCaP cells by inhibition of caspase-9 (A) LNCaP and DU145 cells were treated with ABT-263 (2.5 μM for LNCaP and 10 μM for DU145) in the presence of either or both the caspase-9 and caspase-8 inhibitors (20 μM). After 24 h, cells were stained with FITC-conjugated Annexin V and PI, and flow cytometry was performed. The results are shown as the means + SD of three samples. * P
    Figure Legend Snippet: Induction of caspase-8-dependent ABT-263-induced apoptosis of LNCaP cells by inhibition of caspase-9 (A) LNCaP and DU145 cells were treated with ABT-263 (2.5 μM for LNCaP and 10 μM for DU145) in the presence of either or both the caspase-9 and caspase-8 inhibitors (20 μM). After 24 h, cells were stained with FITC-conjugated Annexin V and PI, and flow cytometry was performed. The results are shown as the means + SD of three samples. * P

    Techniques Used: Inhibition, Staining, Flow Cytometry, Cytometry

    Analysis of caspase-8-dependent ABT-263-induced apoptosis of PC3 cells under caspase-9 inhibition (A) PC3 cells were treated with a caspase-9 inhibitor (20 μM) and/or ABT-263 (4 μM). After 24 h, cells were harvested and cell lysates assayed for their expression of caspase-3, -8, -9, -2, and c-FLIP by immunoblot. α-tubulin and β-actin were used as loading controls. (B) PC3 cells were treated with both DTX (2.5 nM) and ABT-263 (4 μM) in the presence of the indicated caspase inhibitors. After 24 h, cells were stained with FITC-conjugated Annexin V and PI, and flow cytometry was performed. The numbers represent the percentages of each subset. (C) The results are shown as the means + SD of three samples. * P
    Figure Legend Snippet: Analysis of caspase-8-dependent ABT-263-induced apoptosis of PC3 cells under caspase-9 inhibition (A) PC3 cells were treated with a caspase-9 inhibitor (20 μM) and/or ABT-263 (4 μM). After 24 h, cells were harvested and cell lysates assayed for their expression of caspase-3, -8, -9, -2, and c-FLIP by immunoblot. α-tubulin and β-actin were used as loading controls. (B) PC3 cells were treated with both DTX (2.5 nM) and ABT-263 (4 μM) in the presence of the indicated caspase inhibitors. After 24 h, cells were stained with FITC-conjugated Annexin V and PI, and flow cytometry was performed. The numbers represent the percentages of each subset. (C) The results are shown as the means + SD of three samples. * P

    Techniques Used: Inhibition, Expressing, Staining, Flow Cytometry, Cytometry

    22) Product Images from "The Roles of ROS and Caspases in TRAIL-Induced Apoptosis and Necroptosis in Human Pancreatic Cancer Cells"

    Article Title: The Roles of ROS and Caspases in TRAIL-Induced Apoptosis and Necroptosis in Human Pancreatic Cancer Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0127386

    The roles of caspases in apoptosis and necroptosis of TRAIL-treated cells. (A) MiaPaCa-2 and BxPC-3 cells were cultured with TRAIL (50 ng/mL) for 12 h, and protein expression levels of caspase-3, caspase-8, caspase-9, and caspase-2 were evaluated by immunoblot. α-Tubulin was used as the control. (B) MiaPaCa-2 and BxPC-3 cells were cultured with TRAIL (50 ng/mL) in the presence of a panel of caspase inhibitors (20 μM) for 24 h. After staining with annexin V-FITC/PI, flow cytometric analysis was performed. The numbers represent the proportions of each subset. The percentages of annexin V + cells (C) and annexin V - /PI + cells (D) were determined by flow cytometry. All data points shown represent the mean of three culture wells. * P
    Figure Legend Snippet: The roles of caspases in apoptosis and necroptosis of TRAIL-treated cells. (A) MiaPaCa-2 and BxPC-3 cells were cultured with TRAIL (50 ng/mL) for 12 h, and protein expression levels of caspase-3, caspase-8, caspase-9, and caspase-2 were evaluated by immunoblot. α-Tubulin was used as the control. (B) MiaPaCa-2 and BxPC-3 cells were cultured with TRAIL (50 ng/mL) in the presence of a panel of caspase inhibitors (20 μM) for 24 h. After staining with annexin V-FITC/PI, flow cytometric analysis was performed. The numbers represent the proportions of each subset. The percentages of annexin V + cells (C) and annexin V - /PI + cells (D) were determined by flow cytometry. All data points shown represent the mean of three culture wells. * P

    Techniques Used: Cell Culture, Expressing, Staining, Flow Cytometry, Cytometry

    No crosstalk among caspase-2/-9 and ROS in TRAIL-treated cancer cells. (A) MiaPaCa-2 and BxPC-3 cells were cultured with TRAIL (50 ng/mL) with or without caspase-2 inhibitor (20 μM) for 12 h, and the protein expression levels of caspase-3, -8, and -9 were evaluated by immunoblot. α-Tubulin was used as the control. (B) MiaPaCa-2 and BxPC-3 cells were cultured with TRAIL (50 ng/mL) with or without caspase-9 inhibitor (20 μM) for 12 h, and the protein expression of caspase-2 was evaluated by immunoblot. β-Actin was used as the control. (C) MiaPaCa-2 and BxPC-3 cells were cultured with TRAIL (50 ng/mL) with or without NAC (10 mM) for 12 h, and the protein expression of caspase-8, -2, and -9 was evaluated by immunoblot. β-Actin was used as the control. (D) MiaPaCa-2 and BxPC-3 cells were cultured with TRAIL (50 ng/mL) with the indicated inhibitors (20 μM). As a vehicle control, an equal volume of DMSO was added. After 6 h for MiaPaCa-2 and 12 h for BxPC-3, these cells were cultured with carboxy-H 2 DCFDA for 30 min and examined for ROS levels by flow cytometry. The number represents the mean fluorescence intensity. (E) Data represent the mean of three culture wells. MFI: mean fluorescence intensity. * P
    Figure Legend Snippet: No crosstalk among caspase-2/-9 and ROS in TRAIL-treated cancer cells. (A) MiaPaCa-2 and BxPC-3 cells were cultured with TRAIL (50 ng/mL) with or without caspase-2 inhibitor (20 μM) for 12 h, and the protein expression levels of caspase-3, -8, and -9 were evaluated by immunoblot. α-Tubulin was used as the control. (B) MiaPaCa-2 and BxPC-3 cells were cultured with TRAIL (50 ng/mL) with or without caspase-9 inhibitor (20 μM) for 12 h, and the protein expression of caspase-2 was evaluated by immunoblot. β-Actin was used as the control. (C) MiaPaCa-2 and BxPC-3 cells were cultured with TRAIL (50 ng/mL) with or without NAC (10 mM) for 12 h, and the protein expression of caspase-8, -2, and -9 was evaluated by immunoblot. β-Actin was used as the control. (D) MiaPaCa-2 and BxPC-3 cells were cultured with TRAIL (50 ng/mL) with the indicated inhibitors (20 μM). As a vehicle control, an equal volume of DMSO was added. After 6 h for MiaPaCa-2 and 12 h for BxPC-3, these cells were cultured with carboxy-H 2 DCFDA for 30 min and examined for ROS levels by flow cytometry. The number represents the mean fluorescence intensity. (E) Data represent the mean of three culture wells. MFI: mean fluorescence intensity. * P

    Techniques Used: Cell Culture, Expressing, Flow Cytometry, Cytometry, Fluorescence

    23) Product Images from "Sonic Hedgehog Promotes Tumor Cell Survival by Inhibiting CDON Pro-Apoptotic Activity"

    Article Title: Sonic Hedgehog Promotes Tumor Cell Survival by Inhibiting CDON Pro-Apoptotic Activity

    Journal: PLoS Biology

    doi: 10.1371/journal.pbio.1001623

    Inhibition of SHH triggers cancer cell death via CDON-induced apoptosis. (A) Quantification of SHH expression by Q-RT-PCR in a panel of 45 human colorectal tumors and paired normal tissues. Data are presented as a ratio of SHH expression between tumor and normal tissue for each sample. Overexpression of SHH is considered when more than a 1.5-fold increase in the tumor is observed (indicated by the dashed line). (B) Quantification of SHH expression by Q-RT-PCR in a panel of 105 human tumors. Each tumor sample was compared to a normal paired tissue. For each type of tissue, the percentage of tumors showing an increase in SHH (considered when a more than 1.5-fold increase in expression is observed as compared to the normal tissue) is indicated. (C) Quantification of endogenous secreted SHH by ELISA assay in A549, H522, and H460 cells culture medium. H460 cell line is presented as a negative control. (D) Quantification of endogenous secreted SHH by ELISA assay in the culture medium of A549 cells transfected with scramble or SHH siRNA. (E) CDON immunofluorescence staining of A549 cells transfected with scramble or CDON siRNA using a CDON-specific antibody (in green) as described in the methods section. Nuclei were stained with Hoechst (in blue). (F) Apoptotic cell death induction as measured by TUNEL staining was quantified in A549 and H522 cells transfected with SHH siRNA alone or together with CDON siRNA. (G) Caspase-9 activity was measured in A549 cells 18 h after transfection with SHH siRNA alone or together with CDON siRNA. (H) Nude mice were engrafted with A549 cells by subcutaneous injection of 10 million cells. When the mean tumor volume reached approximately 100 mm 3 , animals were treated twice a week by i.p. injection of scramble or SHH siRNA alone or in combination with CDON siRNA during 4 wk. Mean tumor volume and number of animals for each group are indicated. (I) Representative images of scr siRNA, SHH siRNA, or SHH siRNA+CDON siRNA-treated tumors on day 35. (J) Apoptosis quantification by caspase-3 activity assay on xenografted tumor lysates analyzed after 1 wk of treatment with siRNAs. For (H), error bars indicate s.e.m. Statistical treatment of the data was performed using a two-sided Mann–Whitney test compared to scramble siRNA-treated condition (* p
    Figure Legend Snippet: Inhibition of SHH triggers cancer cell death via CDON-induced apoptosis. (A) Quantification of SHH expression by Q-RT-PCR in a panel of 45 human colorectal tumors and paired normal tissues. Data are presented as a ratio of SHH expression between tumor and normal tissue for each sample. Overexpression of SHH is considered when more than a 1.5-fold increase in the tumor is observed (indicated by the dashed line). (B) Quantification of SHH expression by Q-RT-PCR in a panel of 105 human tumors. Each tumor sample was compared to a normal paired tissue. For each type of tissue, the percentage of tumors showing an increase in SHH (considered when a more than 1.5-fold increase in expression is observed as compared to the normal tissue) is indicated. (C) Quantification of endogenous secreted SHH by ELISA assay in A549, H522, and H460 cells culture medium. H460 cell line is presented as a negative control. (D) Quantification of endogenous secreted SHH by ELISA assay in the culture medium of A549 cells transfected with scramble or SHH siRNA. (E) CDON immunofluorescence staining of A549 cells transfected with scramble or CDON siRNA using a CDON-specific antibody (in green) as described in the methods section. Nuclei were stained with Hoechst (in blue). (F) Apoptotic cell death induction as measured by TUNEL staining was quantified in A549 and H522 cells transfected with SHH siRNA alone or together with CDON siRNA. (G) Caspase-9 activity was measured in A549 cells 18 h after transfection with SHH siRNA alone or together with CDON siRNA. (H) Nude mice were engrafted with A549 cells by subcutaneous injection of 10 million cells. When the mean tumor volume reached approximately 100 mm 3 , animals were treated twice a week by i.p. injection of scramble or SHH siRNA alone or in combination with CDON siRNA during 4 wk. Mean tumor volume and number of animals for each group are indicated. (I) Representative images of scr siRNA, SHH siRNA, or SHH siRNA+CDON siRNA-treated tumors on day 35. (J) Apoptosis quantification by caspase-3 activity assay on xenografted tumor lysates analyzed after 1 wk of treatment with siRNAs. For (H), error bars indicate s.e.m. Statistical treatment of the data was performed using a two-sided Mann–Whitney test compared to scramble siRNA-treated condition (* p

    Techniques Used: Inhibition, Expressing, Reverse Transcription Polymerase Chain Reaction, Over Expression, Enzyme-linked Immunosorbent Assay, Negative Control, Transfection, Immunofluorescence, Staining, TUNEL Assay, Activity Assay, Mouse Assay, Injection, Caspase-3 Activity Assay, MANN-WHITNEY

    CDON triggers apoptosis through CDON proteolytic cleavage and recruitment and activation of caspase-9. (A) In vitro –translated CDON intracellular domain (CDON-IC) wild-type (wt) or mutated on one (left panel) or two (right panel) aspartic acid residues were incubated in the absence or in the presence of recombinant purified active caspase-3. Autoradiographs show the cleavage by caspase-3 of CDON-IC wt, whereas CDON-IC–D1153N was weakly cleaved and CDON-IC–D1153N–D1164N was almost completely resistant to cleavage. The band appearing in the CDON D1164N–D1153N probably represents a cryptic site. (B) Schematic representation of CDON and its different mutant constructs. CDON-main (D1153) and secondary (D1178) caspase cleavage sites are shown. (C–D) Apoptotic cell death induction as measured by caspase-3 activity (C) was quantified in HEK293T cells transfected with wild-type (CDON) full-length or mutated full-length CDON expression plasmids and by TUNEL (D) staining was quantified in HEK293T cells transfected with wild-type (CDON) full-length or mutated full-length CDON expression plasmids. Quantifications of TUNEL positive cells (orange) and nuclei (Hoechst in blue) are shown. (E) Apoptotic cell death induction as measured by caspase-3 activity was quantified in HEK293T cells transfected with constructs encoding full-length CDON or the CDON hypothetical fragments resulting from its cleavage by caspase at D1153 (CDON1–1153 and CDON 1154–1250). (F) HEK293T cells were transfected with constructs encoding CDON and/or caspase-9 and cell lysates were subjected to immunoprecipitation with a CDON-specific antibody. CDON and caspase-9 proteins were detected by Western blot in immunoprecipitated and input fractions. (G) Caspase-9 activity was quantified in HEK293T cells transfected with wild-type (CDON) full-length or mutated full-length CDON (D1153N) expression plasmids. For (C–F) and (G), data are means of at least three independent assays. Error bars indicate s.d.
    Figure Legend Snippet: CDON triggers apoptosis through CDON proteolytic cleavage and recruitment and activation of caspase-9. (A) In vitro –translated CDON intracellular domain (CDON-IC) wild-type (wt) or mutated on one (left panel) or two (right panel) aspartic acid residues were incubated in the absence or in the presence of recombinant purified active caspase-3. Autoradiographs show the cleavage by caspase-3 of CDON-IC wt, whereas CDON-IC–D1153N was weakly cleaved and CDON-IC–D1153N–D1164N was almost completely resistant to cleavage. The band appearing in the CDON D1164N–D1153N probably represents a cryptic site. (B) Schematic representation of CDON and its different mutant constructs. CDON-main (D1153) and secondary (D1178) caspase cleavage sites are shown. (C–D) Apoptotic cell death induction as measured by caspase-3 activity (C) was quantified in HEK293T cells transfected with wild-type (CDON) full-length or mutated full-length CDON expression plasmids and by TUNEL (D) staining was quantified in HEK293T cells transfected with wild-type (CDON) full-length or mutated full-length CDON expression plasmids. Quantifications of TUNEL positive cells (orange) and nuclei (Hoechst in blue) are shown. (E) Apoptotic cell death induction as measured by caspase-3 activity was quantified in HEK293T cells transfected with constructs encoding full-length CDON or the CDON hypothetical fragments resulting from its cleavage by caspase at D1153 (CDON1–1153 and CDON 1154–1250). (F) HEK293T cells were transfected with constructs encoding CDON and/or caspase-9 and cell lysates were subjected to immunoprecipitation with a CDON-specific antibody. CDON and caspase-9 proteins were detected by Western blot in immunoprecipitated and input fractions. (G) Caspase-9 activity was quantified in HEK293T cells transfected with wild-type (CDON) full-length or mutated full-length CDON (D1153N) expression plasmids. For (C–F) and (G), data are means of at least three independent assays. Error bars indicate s.d.

    Techniques Used: Activation Assay, In Vitro, Incubation, Recombinant, Purification, Mutagenesis, Construct, Activity Assay, Transfection, Expressing, TUNEL Assay, Staining, Immunoprecipitation, Western Blot

    24) Product Images from "Hyperthermia enhances mapatumumab-induced apoptotic death through ubiquitin-mediated degradation of cellular FLIP(long) in human colon cancer cells"

    Article Title: Hyperthermia enhances mapatumumab-induced apoptotic death through ubiquitin-mediated degradation of cellular FLIP(long) in human colon cancer cells

    Journal: Cell Death & Disease

    doi: 10.1038/cddis.2013.104

    c-FLIP L was responsible for the synergistic effect of Mapa and hyperthermia. ( a ) CX-1 cells were transfected with pCR3.V64-Met-Flag-FLIPL and stable clones were selected with G418 (500 μ g/ml). ( b ) Control plasmid or pCR3.V64-Met-Flag-FLIPL stably transfected cells from a pool of clone 1#, 8# and 11# were heated at 42 °C for 1 h in the presence or absence of 500 ng/ml Mapa, and then incubated at 37 °C for 3 h. The level of c-FLIP L and the cleavage of caspase 8, caspase 9 and PARP were detected by immunoblotting. Actin was used as loading control. ( c ) CX-1 cells were transfected with nonsense sequence (control) or FLIP siRNA-targeting FLIP mRNA. After 48 h, cells were heated at 42 °C for 1 h in the presence or absence of 100 ng/ml Mapa, and then incubated for 3 h. The levels of c-FLIP L and PARP were detected by immunoblotting. Actin was used as a loading control
    Figure Legend Snippet: c-FLIP L was responsible for the synergistic effect of Mapa and hyperthermia. ( a ) CX-1 cells were transfected with pCR3.V64-Met-Flag-FLIPL and stable clones were selected with G418 (500 μ g/ml). ( b ) Control plasmid or pCR3.V64-Met-Flag-FLIPL stably transfected cells from a pool of clone 1#, 8# and 11# were heated at 42 °C for 1 h in the presence or absence of 500 ng/ml Mapa, and then incubated at 37 °C for 3 h. The level of c-FLIP L and the cleavage of caspase 8, caspase 9 and PARP were detected by immunoblotting. Actin was used as loading control. ( c ) CX-1 cells were transfected with nonsense sequence (control) or FLIP siRNA-targeting FLIP mRNA. After 48 h, cells were heated at 42 °C for 1 h in the presence or absence of 100 ng/ml Mapa, and then incubated for 3 h. The levels of c-FLIP L and PARP were detected by immunoblotting. Actin was used as a loading control

    Techniques Used: Transfection, Clone Assay, Plasmid Preparation, Stable Transfection, Incubation, Sequencing

    25) Product Images from "Induction of Lysosome‐associated Protein Transmembrane 4 Beta via Sulfatase 2 Enhances Autophagic Flux in Liver Cancer Cells"

    Article Title: Induction of Lysosome‐associated Protein Transmembrane 4 Beta via Sulfatase 2 Enhances Autophagic Flux in Liver Cancer Cells

    Journal: Hepatology Communications

    doi: 10.1002/hep4.1429

    SULF2 is associated with expression of lysosome‐related proteins. Western blot analyses show protein expressions of LAMP1, cathepsin D, and cathepsin B (left column) according to SULF2 status in Huh7 and Hep3B cells. Common markers for apoptosis (right column) are also assessed. Abbreviations: Cas‐8, caspase‐8; Cas‐9, caspase‐9; CTSB, cathepsin B; CTSD, cathepsin D; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; LAMP1, lysosome‐associated membrane protein 1; SULF2, sulfatase 2.
    Figure Legend Snippet: SULF2 is associated with expression of lysosome‐related proteins. Western blot analyses show protein expressions of LAMP1, cathepsin D, and cathepsin B (left column) according to SULF2 status in Huh7 and Hep3B cells. Common markers for apoptosis (right column) are also assessed. Abbreviations: Cas‐8, caspase‐8; Cas‐9, caspase‐9; CTSB, cathepsin B; CTSD, cathepsin D; GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; LAMP1, lysosome‐associated membrane protein 1; SULF2, sulfatase 2.

    Techniques Used: Expressing, Western Blot

    26) Product Images from "Bishonokiol A Induces Multiple Cell Death in Human Breast Cancer MCF-7 Cells"

    Article Title: Bishonokiol A Induces Multiple Cell Death in Human Breast Cancer MCF-7 Cells

    Journal: Asian Pacific Journal of Cancer Prevention : APJCP

    doi: 10.31557/APJCP.2020.21.4.1073

    BHNKA Induced Apoptosis in Human Breast Cancer MCF-7 Cells. (A), Cell death was analyzed using flow cytometric with PI staining exposed to various concentration of BHNKA for 24 h. (B), MCF-7 cells were treatment with various concentration of BHNKA for 24 h, subjected to DAPI staining and visualized by fluorescence microscopy. Red arrowheads indicated chromatin concentration and nuclear pyknosis in massive cells. (C), Electron microscopy were used to analyze morphological characteristics of apoptosis. (D), The expression of caspase-9, Bax and Bcl-2 proteins were analyzed by western blotting. GAPDH served as loading control. (E), Cell viability treated with DMSO, z-VAD (20 μM), BHNKA (40 μM), and BHNKA pre-treatment with z-VAD were analyzed by MTT assay. * P
    Figure Legend Snippet: BHNKA Induced Apoptosis in Human Breast Cancer MCF-7 Cells. (A), Cell death was analyzed using flow cytometric with PI staining exposed to various concentration of BHNKA for 24 h. (B), MCF-7 cells were treatment with various concentration of BHNKA for 24 h, subjected to DAPI staining and visualized by fluorescence microscopy. Red arrowheads indicated chromatin concentration and nuclear pyknosis in massive cells. (C), Electron microscopy were used to analyze morphological characteristics of apoptosis. (D), The expression of caspase-9, Bax and Bcl-2 proteins were analyzed by western blotting. GAPDH served as loading control. (E), Cell viability treated with DMSO, z-VAD (20 μM), BHNKA (40 μM), and BHNKA pre-treatment with z-VAD were analyzed by MTT assay. * P

    Techniques Used: Staining, Concentration Assay, Fluorescence, Microscopy, Electron Microscopy, Expressing, Western Blot, MTT Assay

    27) Product Images from "Composite fatty acid ether amides suppress growth of liver cancer cells in vitro and in an in vivo allograft mouse model"

    Article Title: Composite fatty acid ether amides suppress growth of liver cancer cells in vitro and in an in vivo allograft mouse model

    Journal: Cellular oncology (Dordrecht)

    doi: 10.1007/s13402-013-0132-x

    Apoptosis induction and beta-spectrin expression in Huh7 and 1MEA cells. a . Expression of survivin, LC3-I, LC3-II, cleaved caspase 7,cleaved caspase 9 and cleaved PARP in Huh7 cells treated with or without AIP-1and AIP-2. b Expression of survivin, LC3-I,
    Figure Legend Snippet: Apoptosis induction and beta-spectrin expression in Huh7 and 1MEA cells. a . Expression of survivin, LC3-I, LC3-II, cleaved caspase 7,cleaved caspase 9 and cleaved PARP in Huh7 cells treated with or without AIP-1and AIP-2. b Expression of survivin, LC3-I,

    Techniques Used: Expressing

    28) Product Images from "Daphnoretin-induced apoptosis in HeLa cells: a possible mitochondria-dependent pathway"

    Article Title: Daphnoretin-induced apoptosis in HeLa cells: a possible mitochondria-dependent pathway

    Journal: Cytotechnology

    doi: 10.1007/s10616-013-9536-8

    Daphnoretin induces both caspase-3 and caspase-9 activity. After treatment with daphnoretin (0.1, 1, 10, 50 μg/ml) or carboplatin (500 μg/ml) for 24 h, caspase activity of HeLa cells was detected by a colorimetric
    Figure Legend Snippet: Daphnoretin induces both caspase-3 and caspase-9 activity. After treatment with daphnoretin (0.1, 1, 10, 50 μg/ml) or carboplatin (500 μg/ml) for 24 h, caspase activity of HeLa cells was detected by a colorimetric

    Techniques Used: Activity Assay

    29) Product Images from "Antitumor effect of pyrrolo-1,5-benzoxazepine-15 and its synergistic effect with Oxaliplatin and 5-FU in colorectal cancer cells"

    Article Title: Antitumor effect of pyrrolo-1,5-benzoxazepine-15 and its synergistic effect with Oxaliplatin and 5-FU in colorectal cancer cells

    Journal: Cancer Biology & Therapy

    doi: 10.1080/15384047.2015.1078028

    PBOX-15 affects both the apoptotic cascade and MAPKs pathway. ( A and B ) Lysates from DLD-1 cells, untreated (−) or treated with PBOX-15 for the indicated time points at the concentrations of 100 nM and 1 µM, were immunoblotted with anti-caspase-9, anti-caspase-3, anti-PARP (total and cleaved forms) and anti-pH2AX antibodies (panel A ), and with anti-ERK or anti-pERK antibodies (panel B ). To confirm equal loading the filters were stripped and reprobed with anti-GAPDH antibody. ( C ) Lysates from DLD-1 cells untreated (−) or treated (+) with PBOX-15 (1 µM) for the indicated incubation times, were immunoblotted with anti-JNK and anti-pJNK antibodies. Blots shown are representative of at least 3 independent experiments.
    Figure Legend Snippet: PBOX-15 affects both the apoptotic cascade and MAPKs pathway. ( A and B ) Lysates from DLD-1 cells, untreated (−) or treated with PBOX-15 for the indicated time points at the concentrations of 100 nM and 1 µM, were immunoblotted with anti-caspase-9, anti-caspase-3, anti-PARP (total and cleaved forms) and anti-pH2AX antibodies (panel A ), and with anti-ERK or anti-pERK antibodies (panel B ). To confirm equal loading the filters were stripped and reprobed with anti-GAPDH antibody. ( C ) Lysates from DLD-1 cells untreated (−) or treated (+) with PBOX-15 (1 µM) for the indicated incubation times, were immunoblotted with anti-JNK and anti-pJNK antibodies. Blots shown are representative of at least 3 independent experiments.

    Techniques Used: Incubation

    30) Product Images from "Adenosine induces intrinsic apoptosis via the PI3K/Akt/mTOR signaling pathway in human pharyngeal squamous carcinoma FaDu cells"

    Article Title: Adenosine induces intrinsic apoptosis via the PI3K/Akt/mTOR signaling pathway in human pharyngeal squamous carcinoma FaDu cells

    Journal: Oncology Letters

    doi: 10.3892/ol.2018.8089

    Bax, Bcl-2, and cytochrome c mediate adenosine-induced apoptosis via the mitochondrial intrinsic pathway. FaDu cells were treated with 3 mM adenosine for 24 h, prior to collection of whole cell lysates. Samples were separated using (8–15%) SDS-PAGE, and resolved by incubation with primary antibodies against (A) Bax, Bcl-2, cytochrome c, and (B) caspase-9, caspase-3, and PARP. β-actin was used as an internal control for the western blot analysis. Data are representative of three experiments that produced similar results. Sample bands (n=3) were densitometrically evaluated. Bcl-2, B-cell lymphoma 2; Bax, Bcl-associated X; PARP, poly(ADP-ribose) polymerase.
    Figure Legend Snippet: Bax, Bcl-2, and cytochrome c mediate adenosine-induced apoptosis via the mitochondrial intrinsic pathway. FaDu cells were treated with 3 mM adenosine for 24 h, prior to collection of whole cell lysates. Samples were separated using (8–15%) SDS-PAGE, and resolved by incubation with primary antibodies against (A) Bax, Bcl-2, cytochrome c, and (B) caspase-9, caspase-3, and PARP. β-actin was used as an internal control for the western blot analysis. Data are representative of three experiments that produced similar results. Sample bands (n=3) were densitometrically evaluated. Bcl-2, B-cell lymphoma 2; Bax, Bcl-associated X; PARP, poly(ADP-ribose) polymerase.

    Techniques Used: SDS Page, Incubation, Western Blot, Produced

    31) Product Images from "Extracts of Artocarpus communis Induce Mitochondria-Associated Apoptosis via Pro-oxidative Activity in Human Glioblastoma Cells"

    Article Title: Extracts of Artocarpus communis Induce Mitochondria-Associated Apoptosis via Pro-oxidative Activity in Human Glioblastoma Cells

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2018.00411

    Artocarpin inhibits xenograft tumor growth in SCID mice. (A,B) Mice were subcutaneously injected with U87 cells. When the tumors grew to 100 mm 3 , the mice were administered artocarpin (2 mg kg -1 ) or vehicle once daily for 3 weeks. Tumor volume was determined at various times after implantation ( n = 8–10). (C) Mouse body weights were measured at various times after tumor implantation. (D) Western blot analyses of Bax, Bcl-2, cleaved caspase-3, cleaved caspase-9, cleaved PARP, phospho-ERK1/2, and phospho-Akt levels in tumors with- and without artocarpin treatment. ∗ P
    Figure Legend Snippet: Artocarpin inhibits xenograft tumor growth in SCID mice. (A,B) Mice were subcutaneously injected with U87 cells. When the tumors grew to 100 mm 3 , the mice were administered artocarpin (2 mg kg -1 ) or vehicle once daily for 3 weeks. Tumor volume was determined at various times after implantation ( n = 8–10). (C) Mouse body weights were measured at various times after tumor implantation. (D) Western blot analyses of Bax, Bcl-2, cleaved caspase-3, cleaved caspase-9, cleaved PARP, phospho-ERK1/2, and phospho-Akt levels in tumors with- and without artocarpin treatment. ∗ P

    Techniques Used: Mouse Assay, Injection, Tumor Implantation, Western Blot

    Artocarpin inhibits xenograft tumor growth in SCID mice. (A,B) Mice were subcutaneously injected with U118 cells. When the tumors grew to 100 mm 3 , the mice were administered artocarpin (2 mg kg -1 ) or vehicle once daily for 3 weeks. Tumor volume was determined at various times after implantation ( n = 8–10). (C) Mouse body weights were measured at various times after tumor implantation. (D) Western blot analyses for Bax, Bcl-2, cleaved caspase-3, cleaved caspase-9, cleaved PARP, phospho-ERK1/2, and phospho-Akt levels in tumors with- and without artocarpin treatment. ∗ P
    Figure Legend Snippet: Artocarpin inhibits xenograft tumor growth in SCID mice. (A,B) Mice were subcutaneously injected with U118 cells. When the tumors grew to 100 mm 3 , the mice were administered artocarpin (2 mg kg -1 ) or vehicle once daily for 3 weeks. Tumor volume was determined at various times after implantation ( n = 8–10). (C) Mouse body weights were measured at various times after tumor implantation. (D) Western blot analyses for Bax, Bcl-2, cleaved caspase-3, cleaved caspase-9, cleaved PARP, phospho-ERK1/2, and phospho-Akt levels in tumors with- and without artocarpin treatment. ∗ P

    Techniques Used: Mouse Assay, Injection, Tumor Implantation, Western Blot

    32) Product Images from "Suppression of the NF-κB signaling pathway in colon cancer cells by the natural compound Riccardin D from Dumortierahirsute"

    Article Title: Suppression of the NF-κB signaling pathway in colon cancer cells by the natural compound Riccardin D from Dumortierahirsute

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2018.8617

    Riccardin D induces HT-29 cell apoptosis via a caspase-dependent pathway. (A) Morphology was observed in HT-29 cells with Riccardin D treatment induced apoptosis. Cells were treated with 0 (control), 5, 10 and 20 µM Riccardin D treated. Arrows indicate apoptotic characteristics in treated cells (scale bar: 50 µm). (B) Riccardin D induced a reduction in the mitochondrial membrane potential of HT-29 cells. A decrease in the red fluorescence ratio indicated a shift associated with a reduction in mitochondrial depolarization following treatment with 5–20 µM Riccardin D (scale bar: 50 µm). (C) Riccardin D triggers HT-29 cell apoptosis via the caspase-dependent pathway. The protein expression levels of cleaved caspase-3, caspase-9 and PARP in HT-29 cells were increased by Riccardin D treatment. In addition, Bcl-2 protein expression was decreased and Bax expression was increased in cancer cells treated with 20 µM Riccardin D (n=3 repeated experiments). PARP, poly (adenosine diphosphate-ribose) polymerase; Bcl-2, B-cell lymphoma 2; BAX, Bcl-2-associated X protein.
    Figure Legend Snippet: Riccardin D induces HT-29 cell apoptosis via a caspase-dependent pathway. (A) Morphology was observed in HT-29 cells with Riccardin D treatment induced apoptosis. Cells were treated with 0 (control), 5, 10 and 20 µM Riccardin D treated. Arrows indicate apoptotic characteristics in treated cells (scale bar: 50 µm). (B) Riccardin D induced a reduction in the mitochondrial membrane potential of HT-29 cells. A decrease in the red fluorescence ratio indicated a shift associated with a reduction in mitochondrial depolarization following treatment with 5–20 µM Riccardin D (scale bar: 50 µm). (C) Riccardin D triggers HT-29 cell apoptosis via the caspase-dependent pathway. The protein expression levels of cleaved caspase-3, caspase-9 and PARP in HT-29 cells were increased by Riccardin D treatment. In addition, Bcl-2 protein expression was decreased and Bax expression was increased in cancer cells treated with 20 µM Riccardin D (n=3 repeated experiments). PARP, poly (adenosine diphosphate-ribose) polymerase; Bcl-2, B-cell lymphoma 2; BAX, Bcl-2-associated X protein.

    Techniques Used: Fluorescence, Expressing

    33) Product Images from "Lup-20(29)-en-3β,28-di-yl-nitrooxy acetate affects MCF-7 proliferation through the crosstalk between apoptosis and autophagy in mitochondria"

    Article Title: Lup-20(29)-en-3β,28-di-yl-nitrooxy acetate affects MCF-7 proliferation through the crosstalk between apoptosis and autophagy in mitochondria

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-017-0255-5

    NBT induced MCF-7 cell apoptosis through the mitochondrial pathway. a MCF-7 cells were treated with various concentrations of NBT for 24 h, and apoptosis was examined by using annexin V with PI staining with flow cytometry. b Hoechst 33258 staining assay showed that NBT at doses of 6, 12, and 24 μM induced chromatin shrinking of MCF-7 cells (×200, scale bars: 100 μm; ×400, scale bars: 50 μm). c Treatment of 6, 12, and 24 μM NBT influenced caspase-9 activity in MCF-7 cells. d Total cellular extract of MCF-7 cells treated with 6, 12, and 24 μM NBT was prepared and subjected to Western blot by using an antibody against PARP. e JC-1 staining assay determined the ΔΨm in mitochondria of MCF-7 cells treated with 6, 12, and 24 μM NBT. f Total cellular extract, mitochondrial fraction, and cytosol fractions of MCF-7 cells treated with 6, 12, and 24 μM NBT were prepared and subjected to Western blot by using antibodies against Bcl-2, Bax, and Cyt C (mitochondrial and cytosol fraction). g Total cellular extract of MCF-7 cells treated with 6, 12, and 24 μM NBT was prepared and subjected to Western blot by using antibodies against DRP1, Fis1, and Mfn2. Data represent mean ± SD of three independent experiments ( * P
    Figure Legend Snippet: NBT induced MCF-7 cell apoptosis through the mitochondrial pathway. a MCF-7 cells were treated with various concentrations of NBT for 24 h, and apoptosis was examined by using annexin V with PI staining with flow cytometry. b Hoechst 33258 staining assay showed that NBT at doses of 6, 12, and 24 μM induced chromatin shrinking of MCF-7 cells (×200, scale bars: 100 μm; ×400, scale bars: 50 μm). c Treatment of 6, 12, and 24 μM NBT influenced caspase-9 activity in MCF-7 cells. d Total cellular extract of MCF-7 cells treated with 6, 12, and 24 μM NBT was prepared and subjected to Western blot by using an antibody against PARP. e JC-1 staining assay determined the ΔΨm in mitochondria of MCF-7 cells treated with 6, 12, and 24 μM NBT. f Total cellular extract, mitochondrial fraction, and cytosol fractions of MCF-7 cells treated with 6, 12, and 24 μM NBT were prepared and subjected to Western blot by using antibodies against Bcl-2, Bax, and Cyt C (mitochondrial and cytosol fraction). g Total cellular extract of MCF-7 cells treated with 6, 12, and 24 μM NBT was prepared and subjected to Western blot by using antibodies against DRP1, Fis1, and Mfn2. Data represent mean ± SD of three independent experiments ( * P

    Techniques Used: Staining, Flow Cytometry, Cytometry, Activity Assay, Western Blot

    34) Product Images from "EGR1 interacts with TBX2 and functions as a tumor suppressor in rhabdomyosarcoma"

    Article Title: EGR1 interacts with TBX2 and functions as a tumor suppressor in rhabdomyosarcoma

    Journal: Oncotarget

    doi: 10.18632/oncotarget.24726

    EGR1 over expression induces intrinsic cell apoptosis ( A ) RH30 cells over expressing EGR1 or vector control were subject to differentiation conditions for 4 days and apoptosis was detected by TUNEL assay. ( B ) Caspase 8 cleavage is not affected by EGR1. Cells as described in A were assayed by western blot analysis. ( C ) Caspase 9 cleavage is induced by EGR1, assayed as in (B). Single asterisk indicates full length protein; double asterisks indicate cleaved products. ( D ) Caspase 3 and p-p38 are induced by EGR1, assayed as in B. BAX1 is upregulated by EGR as assayed by qRT-PCR ( E ) and western blot assay ( F ). Error Bars, S.D. and *** P > 0.001 vs. vector ( G ) Phosphorylated BAD (pBAD) is downregulated in response to EGR1.
    Figure Legend Snippet: EGR1 over expression induces intrinsic cell apoptosis ( A ) RH30 cells over expressing EGR1 or vector control were subject to differentiation conditions for 4 days and apoptosis was detected by TUNEL assay. ( B ) Caspase 8 cleavage is not affected by EGR1. Cells as described in A were assayed by western blot analysis. ( C ) Caspase 9 cleavage is induced by EGR1, assayed as in (B). Single asterisk indicates full length protein; double asterisks indicate cleaved products. ( D ) Caspase 3 and p-p38 are induced by EGR1, assayed as in B. BAX1 is upregulated by EGR as assayed by qRT-PCR ( E ) and western blot assay ( F ). Error Bars, S.D. and *** P > 0.001 vs. vector ( G ) Phosphorylated BAD (pBAD) is downregulated in response to EGR1.

    Techniques Used: Over Expression, Expressing, Plasmid Preparation, TUNEL Assay, Western Blot, Quantitative RT-PCR

    35) Product Images from "Receptor Interactive Protein Kinase 3 Promotes Cisplatin-Triggered Necrosis in Apoptosis-Resistant Esophageal Squamous Cell Carcinoma Cells"

    Article Title: Receptor Interactive Protein Kinase 3 Promotes Cisplatin-Triggered Necrosis in Apoptosis-Resistant Esophageal Squamous Cell Carcinoma Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0100127

    Clustering display of the microarray data after cisplatin treatment. (A) RNA was extracted using an RNA isolation kit for microarray analysis after cisplatin treatment for 6 h. The up-regulated genes were functionally classified based on their biological process using the DAVID functional annotation clustering tool. (B) The mRNA levels of cell-death-associated genes in KYSE140 cells treated with 10 µM cisplatin for 6 h were determined by real-time RT-PCR. The data represent the mean ± SEM of relative mRNA levels versus untreated cells. (C) Protein levels of FOS, RIPK3, MLKL, Cytochrome c, survivin and caspase-9 in KYSE140 cells after treatment with 10 µM cisplatin for the indicated time points were analyzed by western blotting. (D) The mRNA levels of cell-death-associated genes in KYSE140 and EC0156 cells treated with 10 µM cisplatin for 6 h were determined by real-time PCR. **, P
    Figure Legend Snippet: Clustering display of the microarray data after cisplatin treatment. (A) RNA was extracted using an RNA isolation kit for microarray analysis after cisplatin treatment for 6 h. The up-regulated genes were functionally classified based on their biological process using the DAVID functional annotation clustering tool. (B) The mRNA levels of cell-death-associated genes in KYSE140 cells treated with 10 µM cisplatin for 6 h were determined by real-time RT-PCR. The data represent the mean ± SEM of relative mRNA levels versus untreated cells. (C) Protein levels of FOS, RIPK3, MLKL, Cytochrome c, survivin and caspase-9 in KYSE140 cells after treatment with 10 µM cisplatin for the indicated time points were analyzed by western blotting. (D) The mRNA levels of cell-death-associated genes in KYSE140 and EC0156 cells treated with 10 µM cisplatin for 6 h were determined by real-time PCR. **, P

    Techniques Used: Microarray, Isolation, Functional Assay, Quantitative RT-PCR, Western Blot, Real-time Polymerase Chain Reaction

    36) Product Images from "OGG1 Involvement in High Glucose-Mediated Enhancement of Bupivacaine-Induced Oxidative DNA Damage in SH-SY5Y Cells"

    Article Title: OGG1 Involvement in High Glucose-Mediated Enhancement of Bupivacaine-Induced Oxidative DNA Damage in SH-SY5Y Cells

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2015/683197

    The effect of high glucose, bupivacaine, or NAC on OGG1 and cleaved caspase-9 protein expression as detected by western blot. Con: SH-SY5Y cells of group control. HG: SH-SY5Y cells were exposed to 50 mM glucose for 8 d. Bup: SH-SY5Y cells were treated with 1.5 mM bupivacaine for 24 h. HG + Bup: SH-SY5Y cells were incubated with 50 mM glucose for 8 d and then treated with 1.5 mM bupivacaine for 24 h. HG + Bup + NAC: cells treated with 50 mM glucose for 8 d and then pretreated with 5 mM NAC for 6 h prior to 1.5 mM bupivacaine exposure for 24 h. (a) The effect of high glucose on OGG1 and cleaved caspase-9 protein expression induced by bupivacaine. (b) The effect of NAC on OGG1 and cleaved caspase-9 protein expression induced by high glucose and bupivacaine. Data are presented as mean ± SD ( n = 3) ∗ P
    Figure Legend Snippet: The effect of high glucose, bupivacaine, or NAC on OGG1 and cleaved caspase-9 protein expression as detected by western blot. Con: SH-SY5Y cells of group control. HG: SH-SY5Y cells were exposed to 50 mM glucose for 8 d. Bup: SH-SY5Y cells were treated with 1.5 mM bupivacaine for 24 h. HG + Bup: SH-SY5Y cells were incubated with 50 mM glucose for 8 d and then treated with 1.5 mM bupivacaine for 24 h. HG + Bup + NAC: cells treated with 50 mM glucose for 8 d and then pretreated with 5 mM NAC for 6 h prior to 1.5 mM bupivacaine exposure for 24 h. (a) The effect of high glucose on OGG1 and cleaved caspase-9 protein expression induced by bupivacaine. (b) The effect of NAC on OGG1 and cleaved caspase-9 protein expression induced by high glucose and bupivacaine. Data are presented as mean ± SD ( n = 3) ∗ P

    Techniques Used: Expressing, Western Blot, Incubation

    37) Product Images from "Quercetin reverses the doxorubicin resistance of prostate cancer cells by downregulating the expression of c-met"

    Article Title: Quercetin reverses the doxorubicin resistance of prostate cancer cells by downregulating the expression of c-met

    Journal: Oncology Letters

    doi: 10.3892/ol.2017.7561

    Quercetin promoted doxorubicin-induced apoptosis via the mitochondrial pathway. (A) The mitochondrial membrane potential of PC3/R cells treated with quercetin (10 µM) and doxorubicin (2 µg/ml) was measured by JC-1 staining and flow cytometry. (B) The generation of ROS in PC3/R cells treated with quercetin (10 µM) and doxorubicin (2 µg/ml) was measured by DHE staining and flow cytometry. (C) Western blot analysis was performed to evaluate the cleavage of caspase-9 and caspase-3 in the PC3/R cells treated with quercetin (10 µM) and doxorubicin (2 µg/ml). PC3/R, prostate cancer 3/resistant; ROS, reactive oxygen species; DHE, dihydroethidium.
    Figure Legend Snippet: Quercetin promoted doxorubicin-induced apoptosis via the mitochondrial pathway. (A) The mitochondrial membrane potential of PC3/R cells treated with quercetin (10 µM) and doxorubicin (2 µg/ml) was measured by JC-1 staining and flow cytometry. (B) The generation of ROS in PC3/R cells treated with quercetin (10 µM) and doxorubicin (2 µg/ml) was measured by DHE staining and flow cytometry. (C) Western blot analysis was performed to evaluate the cleavage of caspase-9 and caspase-3 in the PC3/R cells treated with quercetin (10 µM) and doxorubicin (2 µg/ml). PC3/R, prostate cancer 3/resistant; ROS, reactive oxygen species; DHE, dihydroethidium.

    Techniques Used: Staining, Flow Cytometry, Cytometry, Western Blot

    38) Product Images from "ATP-competitive Plk1 inhibitors induce caspase 3-mediated Plk1 cleavage and activation in hematopoietic cell lines"

    Article Title: ATP-competitive Plk1 inhibitors induce caspase 3-mediated Plk1 cleavage and activation in hematopoietic cell lines

    Journal: Oncotarget

    doi: 10.18632/oncotarget.23650

    Caspase 3 is responsible for Plk1 cleavage ( A ) K562 cells were incubated for 48 h with increasing concentrations of BI-2536 in presence of 50 µM qVD. Expression and cleavage of Plk1 were analysed by Western blot. ( B ) Plk1 was transcribed/translated in a TNT assay and the generated Plk1 protein is incubated with indicated recombinant caspases (C2, C3 and C6 to C9) in the presence or absence of 50 μM qVD. Expression and cleavage of Plk1 were analyzed by Western blot. ( C and D ) After a 5 day pre-treatment with 1 μg/ml tetracycline to allow knock-down of caspase 3 (K562 shC3, C) or 9 (K562 shC9, D), K562 cells were cultured for 48h with or without 25 nM BI-2536. Expression and cleavage of Plk1, and expression of caspase 3 and caspase 9 were analysed by Western blot. ( E ) Plk1 was transcribed/translated in a TNT assay and the generated Plk1 protein is incubated with recombinant caspase 3 in the presence or absence of 50 μM qVD or 50 nM BI-2536. Expression and cleavage of Plk1 were analysed by Western blot. ( F ) K562 cells were pre-treated with 1 µM Imatinib (ima) for 24 h. Then, cells were treated with 25 nM BI-2536 for 0 to 8 h. Expression and cleavage of Plk1 were analyzed by Western blot. ( G ) Cells were treated with 25 nM BI-2536 for 24 h. Next, cell extracts were separated into cytosol and nuclear-enriched fractions and Plk1 location (Plk1, p-Plk1 and Plk1-CT antibody) was assessed by Western blot in the different fractions. Lamin A and Tubulin served as both loading control and validation of the nuclear and cytoplasmic fractions, respectively. p-Plk1 antibody recognizes the activated (phospho-threonine 210) N-Terminal (NT) epitope and cleavage product NT, and Plk-CT recognizes the C-Terminal epitope and corresponding cleavage product CT. Each panel is representative of at least 3 independent experiments.
    Figure Legend Snippet: Caspase 3 is responsible for Plk1 cleavage ( A ) K562 cells were incubated for 48 h with increasing concentrations of BI-2536 in presence of 50 µM qVD. Expression and cleavage of Plk1 were analysed by Western blot. ( B ) Plk1 was transcribed/translated in a TNT assay and the generated Plk1 protein is incubated with indicated recombinant caspases (C2, C3 and C6 to C9) in the presence or absence of 50 μM qVD. Expression and cleavage of Plk1 were analyzed by Western blot. ( C and D ) After a 5 day pre-treatment with 1 μg/ml tetracycline to allow knock-down of caspase 3 (K562 shC3, C) or 9 (K562 shC9, D), K562 cells were cultured for 48h with or without 25 nM BI-2536. Expression and cleavage of Plk1, and expression of caspase 3 and caspase 9 were analysed by Western blot. ( E ) Plk1 was transcribed/translated in a TNT assay and the generated Plk1 protein is incubated with recombinant caspase 3 in the presence or absence of 50 μM qVD or 50 nM BI-2536. Expression and cleavage of Plk1 were analysed by Western blot. ( F ) K562 cells were pre-treated with 1 µM Imatinib (ima) for 24 h. Then, cells were treated with 25 nM BI-2536 for 0 to 8 h. Expression and cleavage of Plk1 were analyzed by Western blot. ( G ) Cells were treated with 25 nM BI-2536 for 24 h. Next, cell extracts were separated into cytosol and nuclear-enriched fractions and Plk1 location (Plk1, p-Plk1 and Plk1-CT antibody) was assessed by Western blot in the different fractions. Lamin A and Tubulin served as both loading control and validation of the nuclear and cytoplasmic fractions, respectively. p-Plk1 antibody recognizes the activated (phospho-threonine 210) N-Terminal (NT) epitope and cleavage product NT, and Plk-CT recognizes the C-Terminal epitope and corresponding cleavage product CT. Each panel is representative of at least 3 independent experiments.

    Techniques Used: Incubation, Expressing, Western Blot, Generated, Recombinant, Cell Culture

    39) Product Images from "Vinorelbine Potently Induces Placental Cell Death, Does Not Harm Fertility and is a Potential Treatment for Ectopic Pregnancy"

    Article Title: Vinorelbine Potently Induces Placental Cell Death, Does Not Harm Fertility and is a Potential Treatment for Ectopic Pregnancy

    Journal: EBioMedicine

    doi: 10.1016/j.ebiom.2018.01.041

    Vinorelbine causes microtubule condensation, arresting trophoblast cell cycle and leading to activation of apoptosis. (A) Microtubules of JEG3 and HTR-8/SVneo cells after treatment with vinorelbine or control equivalent for 16 h, stained with α-tubulin (green) and DAPI (blue). Scale bar = 10 μm. (B–E) HTR-8/SVneo cell cycle phases after treating with vinorelbine, measured by intracellular staining of propidium iodide-A and FACs. (B) Representative histograms showing control treated cells with the proportions of cells in G 0 /G 1  (green), S (yellow) and phase (purple). Treating with vinorelbine at (C) 1 μM and (d) 10 μM halts cell cycle progression at G 2 /M, reducing the proportion of cells in S and G 0 /G 1  phases. (E) Quantification of relative proportion of cells in G 0 /G 1 , S and G 2 /M phases after vinorelbine treatment. Data are means ± SEM of three independent experiments in triplicate. (F–H) Western blot analysis showing increased protein expression of apoptotic markers after treating JEG3 and HTR-8/SVneo cells with vinorelbine. (F) Representative western blots of JEG3 (left) and HTR-8/SVneo (right) protein expression of Bcl-2, BAX, BNIP3 and caspase 9 and 3. Densitometric analysis of apoptotic markers in (G) JEG3 and (h) HTR-8/SVneo cells. Data are means ± SEM of three independent experiments. *p 
    Figure Legend Snippet: Vinorelbine causes microtubule condensation, arresting trophoblast cell cycle and leading to activation of apoptosis. (A) Microtubules of JEG3 and HTR-8/SVneo cells after treatment with vinorelbine or control equivalent for 16 h, stained with α-tubulin (green) and DAPI (blue). Scale bar = 10 μm. (B–E) HTR-8/SVneo cell cycle phases after treating with vinorelbine, measured by intracellular staining of propidium iodide-A and FACs. (B) Representative histograms showing control treated cells with the proportions of cells in G 0 /G 1 (green), S (yellow) and phase (purple). Treating with vinorelbine at (C) 1 μM and (d) 10 μM halts cell cycle progression at G 2 /M, reducing the proportion of cells in S and G 0 /G 1 phases. (E) Quantification of relative proportion of cells in G 0 /G 1 , S and G 2 /M phases after vinorelbine treatment. Data are means ± SEM of three independent experiments in triplicate. (F–H) Western blot analysis showing increased protein expression of apoptotic markers after treating JEG3 and HTR-8/SVneo cells with vinorelbine. (F) Representative western blots of JEG3 (left) and HTR-8/SVneo (right) protein expression of Bcl-2, BAX, BNIP3 and caspase 9 and 3. Densitometric analysis of apoptotic markers in (G) JEG3 and (h) HTR-8/SVneo cells. Data are means ± SEM of three independent experiments. *p 

    Techniques Used: Activation Assay, Staining, FACS, Western Blot, Expressing

    Vinorelbine does not up-regulate pro-apoptotic molecules in human Fallopian tubes and does not affect subsequent fertility in mice. (A–G) Human fallopian tubes were collected from premenopausal women at the time of hysterectomy and treated with increasing doses of vinorelbine (0.01, 0.1, 1, 100 μM) or equivalent PBS for 24 h. (A) Representative western blot analysis performed for apoptotic markers Bcl-2, BAX, BNIP3, cytochrome  c  and caspase 9 and 3. (B) Densitometric analysis was performed on n = 8 samples for each apoptotic marker. The pro-apoptotic marker Bax was reduced with 0.01 μM vinorelbine. (C-I) Female swiss mice intravenously treated with vinorelbine 5 mg/kg or equivalent PBS, three times over two weeks. Four weeks post final treatment, mice were mated and then culled at embryonic day 17.5, pups, placenta and maternal blood were collected. (C) Litter size, (d) pup weight, (E) placental weight, (F) crown to rump length, (G) fetal placental ratio, (H) maternal serum levels of anti-mullerian hormone and (i) gross fetal morphology did not differ between control and treated groups (n = 7 per a group, no difference in vinorelbine or control treated by un-paired  t -test). Data are means ± SEM. *p 
    Figure Legend Snippet: Vinorelbine does not up-regulate pro-apoptotic molecules in human Fallopian tubes and does not affect subsequent fertility in mice. (A–G) Human fallopian tubes were collected from premenopausal women at the time of hysterectomy and treated with increasing doses of vinorelbine (0.01, 0.1, 1, 100 μM) or equivalent PBS for 24 h. (A) Representative western blot analysis performed for apoptotic markers Bcl-2, BAX, BNIP3, cytochrome c and caspase 9 and 3. (B) Densitometric analysis was performed on n = 8 samples for each apoptotic marker. The pro-apoptotic marker Bax was reduced with 0.01 μM vinorelbine. (C-I) Female swiss mice intravenously treated with vinorelbine 5 mg/kg or equivalent PBS, three times over two weeks. Four weeks post final treatment, mice were mated and then culled at embryonic day 17.5, pups, placenta and maternal blood were collected. (C) Litter size, (d) pup weight, (E) placental weight, (F) crown to rump length, (G) fetal placental ratio, (H) maternal serum levels of anti-mullerian hormone and (i) gross fetal morphology did not differ between control and treated groups (n = 7 per a group, no difference in vinorelbine or control treated by un-paired t -test). Data are means ± SEM. *p 

    Techniques Used: Mouse Assay, Western Blot, Marker

    40) Product Images from "Induction of apoptosis by an oleanolic acid derivative in SMMC-7721 human hepatocellular carcinoma cells is associated with mitochondrial dysfunction"

    Article Title: Induction of apoptosis by an oleanolic acid derivative in SMMC-7721 human hepatocellular carcinoma cells is associated with mitochondrial dysfunction

    Journal: Oncology Letters

    doi: 10.3892/ol.2017.7653

    Oleanolic acid derivative altered the expression levels of apoptotic proteins in SMMC-7721 cells. Cell lysates were prepared from SMMC-7721 cells following incubation with the oleanolic acid derivative (7, 9 or 11 µM) for 48 h. Dimethyl sulfoxide-treated cell lysate was used as the control. Approximately 60 µg of protein per sample was used for immunoblotting of (A) Bcl-2 and Bax, and (B) caspase-9/caspase-3, with β-actin used as the loading control. Bcl-2, B cell lymphoma 2.
    Figure Legend Snippet: Oleanolic acid derivative altered the expression levels of apoptotic proteins in SMMC-7721 cells. Cell lysates were prepared from SMMC-7721 cells following incubation with the oleanolic acid derivative (7, 9 or 11 µM) for 48 h. Dimethyl sulfoxide-treated cell lysate was used as the control. Approximately 60 µg of protein per sample was used for immunoblotting of (A) Bcl-2 and Bax, and (B) caspase-9/caspase-3, with β-actin used as the loading control. Bcl-2, B cell lymphoma 2.

    Techniques Used: Expressing, Incubation

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    Article Snippet: .. For Western blot analysis, the following antibodies were used: rabbit monoclonal anti-PI3K (#4257) anti-Akt (#4691) and anti-pAkt (#4060) and anti-caspase-9 (#9506) were purchased from Cell Signaling Technology (USA). ..

    Incubation:

    Article Title: Acetic acid is an oxidative stressor in gastric cancer cells
    Article Snippet: .. Inc., San Antonio, TX) or anti-rat caspase 9 antibodies (Cell Signaling Technology, Inc., Danvers, MA) were added to Can Get Signal Immunoreaction Enhancer Solution 1; the membranes were then incubated with the solution for 60 min. After the primary antibody solution was aspirated, the membrane was washed three times with 15 ml PBST (phosphate-buffered saline [PBS; Wako] plus 0.1% Tween-20). ..

    other:

    Article Title: Mig-2 attenuates cisplatin-induced apoptosis of human glioma cells in vitro through AKT/JNK and AKT/p38 signaling pathways
    Article Snippet: Antibodies and special reagents Cleaved caspase-3 antibody, cleaved caspase-8 antibody, cleaved caspase-9 antibody, cleaved PARP antibody, Bid antibody, Bax antibody, Bcl-2 antibody, cytochrome c antibody, AKT antibody, p-Akt (Ser473) antibody, ERK5 antibody, p-ERK5 (Thr218/Tyr220) antibody, ERK1/2 antibody, p-ERK1/2 (Thr202/Tyr204) antibody, JNK antibody, p-JNK antibody (Thr183/Tyr185), p38 antibody, p-p38 (Thr180/Tyr182) antibody, GFP antibody and β-actin antibody were purchased from Cell Signaling Technology, Inc (Danvers, MA, USA).

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    Cell Signaling Technology Inc mouse anti human caspase 9
    Western blotting showed that compared with Control, PDOX and DOX reduced ERK phosphorylation, decreased BCL-2 expression, and increased caspase-3 and <t>caspase-9</t> activation.
    Mouse Anti Human Caspase 9, supplied by Cell Signaling Technology Inc, 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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    Cell Signaling Technology Inc anti cleaved caspase 9
    MR affected the expression of apoptosis associated proteins in gastric tissues of CAG rats induced by MNNG combined with irregular diet. The expression levels of Bcl-2, Bax, cleaved caspase-3 and cleaved <t>caspase-9</t> were measured using western blotting. ***P
    Anti Cleaved Caspase 9, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc rabbit anti caspase 9
    BV6 and CIK cells reactivate apoptosis in TE671 cells . TE671 cells (TE) were treated for 4 h with 2.5 μM SMAC mimetic BV6 before BV6 was removed and CIK cells were added for 3 h (lane 6). TE and CIK cells alone (lanes 1 and 2), and after 4 h pre-incubation with 2.5 μM BV6 (lines 3 and 4), as well as TE cells after 3 h treatment with CIK cells (lane 5) served as controls. CIK cells within samples of lanes 5 and 6 were removed, before western blot analysis was performed. Caspase activation and Bid cleavage were analyzed. Cleavage fragments are indicated by arrows. β-Actin served as loading control. CIK cell treatment without (lane 5) or particularly with SMAC mimetic BV6 (lane 6) increased TE killing by activation of the caspase cascade. This was evident from the increased cleavage of caspase-8, -3, and -9 into active cleavage fragments in the presence of BV6 and CIK cells, that is, caspase-8 into active p43 and p41 fragments, caspase-3 into active p17 and p12 fragments and <t>caspase-9</t> into active p37 and p35 fragments. Moreover, Bid was predominately cleaved into tBid, the activated form of Bid, upon BV6 and CIK cell treatment (A) . The addition of zVAD a pan caspase inhibitor reduced CIK cell-mediated killing of TE671 cells by 50% demonstrating that BV6 sensitized target cells towards cytotoxicity of CIK cells by caspase activation (B) .
    Rabbit Anti Caspase 9, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 82 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Western blotting showed that compared with Control, PDOX and DOX reduced ERK phosphorylation, decreased BCL-2 expression, and increased caspase-3 and caspase-9 activation.

    Journal: Journal of Translational Medicine

    Article Title: Targeting therapy of hepatocellular carcinoma with doxorubicin prodrug PDOX increases anti-metastatic effect and reduces toxicity: a preclinical study

    doi: 10.1186/1479-5876-11-192

    Figure Lengend Snippet: Western blotting showed that compared with Control, PDOX and DOX reduced ERK phosphorylation, decreased BCL-2 expression, and increased caspase-3 and caspase-9 activation.

    Article Snippet: Next, they were immunoblotted with a rabbit anti-human p-ERK (4370, CST, MA, USA, dilution 1:1000), rabbit anti- human ERK (4695, CST, MA, USA, dilution 1:1000), rabbit anti-human Bcl-2 (2870, CST, MA, USA, dilution 1:1000), rabbit anti-human caspase-3 (9665, CST, MA, USA, dilution 1:1000), mouse anti-human caspase-9 (9508, CST, MA, USA, dilution 1:1000), and rabbit anti-human β-actin (Santa Cruz, CA, USA, dilution 1:1000) for 3 h. Blots were then incubated with a peroxidase-conjugated sheep anti-rabbit IgG (Santa Cruz, CA, USA, dilution 1:8000) or sheep anti-mouse IgG (Santa Cruz, CA, USA, dilution 1:8000) for 2 h and developed using chemiluminescent detection with a Supersignal West Pico assay kit (Thermo, IL, USA) and autoradiography film.

    Techniques: Western Blot, Expressing, Activation Assay

    MR affected the expression of apoptosis associated proteins in gastric tissues of CAG rats induced by MNNG combined with irregular diet. The expression levels of Bcl-2, Bax, cleaved caspase-3 and cleaved caspase-9 were measured using western blotting. ***P

    Journal: American Journal of Translational Research

    Article Title: Morroniside protects against chronic atrophic gastritis in rat via inhibiting inflammation and apoptosis

    doi:

    Figure Lengend Snippet: MR affected the expression of apoptosis associated proteins in gastric tissues of CAG rats induced by MNNG combined with irregular diet. The expression levels of Bcl-2, Bax, cleaved caspase-3 and cleaved caspase-9 were measured using western blotting. ***P

    Article Snippet: Anti-Bax (cat. no. 14796S), anti-caspase-3 (cat. no. 14220T), anti-cleaved caspase-3 (cat. no. 9661S), anti-caspase-9 (cat. no. 9508T), anti-cleaved caspase-9 (cat. no. 3195S), anti-NF-κB p65 (cat. no. 8242S), anti-p-NF-κB p65 (cat. 3033S), anti-IKKα/β (cat. no. 61294S), anti-p-IKKα/β (cat. no. 2697S), anti-IκB-α (cat. no. 4812S) and anti-GAPDH (cat. no. 5174S) antibodies were obtained from Cell Signaling Technology (Boston, MA, USA).

    Techniques: Expressing, Western Blot

    BV6 and CIK cells reactivate apoptosis in TE671 cells . TE671 cells (TE) were treated for 4 h with 2.5 μM SMAC mimetic BV6 before BV6 was removed and CIK cells were added for 3 h (lane 6). TE and CIK cells alone (lanes 1 and 2), and after 4 h pre-incubation with 2.5 μM BV6 (lines 3 and 4), as well as TE cells after 3 h treatment with CIK cells (lane 5) served as controls. CIK cells within samples of lanes 5 and 6 were removed, before western blot analysis was performed. Caspase activation and Bid cleavage were analyzed. Cleavage fragments are indicated by arrows. β-Actin served as loading control. CIK cell treatment without (lane 5) or particularly with SMAC mimetic BV6 (lane 6) increased TE killing by activation of the caspase cascade. This was evident from the increased cleavage of caspase-8, -3, and -9 into active cleavage fragments in the presence of BV6 and CIK cells, that is, caspase-8 into active p43 and p41 fragments, caspase-3 into active p17 and p12 fragments and caspase-9 into active p37 and p35 fragments. Moreover, Bid was predominately cleaved into tBid, the activated form of Bid, upon BV6 and CIK cell treatment (A) . The addition of zVAD a pan caspase inhibitor reduced CIK cell-mediated killing of TE671 cells by 50% demonstrating that BV6 sensitized target cells towards cytotoxicity of CIK cells by caspase activation (B) .

    Journal: Frontiers in Pediatrics

    Article Title: SMAC Mimetic BV6 Enables Sensitization of Resistant Tumor Cells but also Affects Cytokine-Induced Killer (CIK) Cells: A Potential Challenge for Combination Therapy

    doi: 10.3389/fped.2014.00075

    Figure Lengend Snippet: BV6 and CIK cells reactivate apoptosis in TE671 cells . TE671 cells (TE) were treated for 4 h with 2.5 μM SMAC mimetic BV6 before BV6 was removed and CIK cells were added for 3 h (lane 6). TE and CIK cells alone (lanes 1 and 2), and after 4 h pre-incubation with 2.5 μM BV6 (lines 3 and 4), as well as TE cells after 3 h treatment with CIK cells (lane 5) served as controls. CIK cells within samples of lanes 5 and 6 were removed, before western blot analysis was performed. Caspase activation and Bid cleavage were analyzed. Cleavage fragments are indicated by arrows. β-Actin served as loading control. CIK cell treatment without (lane 5) or particularly with SMAC mimetic BV6 (lane 6) increased TE killing by activation of the caspase cascade. This was evident from the increased cleavage of caspase-8, -3, and -9 into active cleavage fragments in the presence of BV6 and CIK cells, that is, caspase-8 into active p43 and p41 fragments, caspase-3 into active p17 and p12 fragments and caspase-9 into active p37 and p35 fragments. Moreover, Bid was predominately cleaved into tBid, the activated form of Bid, upon BV6 and CIK cell treatment (A) . The addition of zVAD a pan caspase inhibitor reduced CIK cell-mediated killing of TE671 cells by 50% demonstrating that BV6 sensitized target cells towards cytotoxicity of CIK cells by caspase activation (B) .

    Article Snippet: Western blot analysis was performed as described previously using the following antibodies: mouse anti-caspase-8 (1:1000; Alexis Biochemicals, Gruenberg, Germany), rabbit anti-Bid, rabbit anti-caspase-3, and rabbit anti-caspase-9 (1:1000; Cell Signaling, Beverly, MA, USA).

    Techniques: Incubation, Western Blot, Activation Assay

    Dicer knockdown decreases the expression levels of apoptosis-associated proteins induced by cisplatin in ovarian cancer cells. Western blot analyses demonstrated that the expression levels of proteins involved in apoptosis signaling pathways, including P73, P63, P53, caspase-9 and caspase-3, were decreased in shDicer cells when compared with shNC cells following cisplatin treatment in the A2780 and CAOV3 cell lines. Western blot analyses also demonstrated that P21 protein expression was not affected by Dicer downregulation in A2780 and CAOV3 cells, while it was decreased following cisplatin treatment. The results were normalized to the expression of the reference protein β-actin. shRNA, short hairpin RNA; NC, negative control.

    Journal: Molecular Medicine Reports

    Article Title: Dicer affects cisplatin-mediated apoptosis in epithelial ovarian cancer cells

    doi: 10.3892/mmr.2018.9452

    Figure Lengend Snippet: Dicer knockdown decreases the expression levels of apoptosis-associated proteins induced by cisplatin in ovarian cancer cells. Western blot analyses demonstrated that the expression levels of proteins involved in apoptosis signaling pathways, including P73, P63, P53, caspase-9 and caspase-3, were decreased in shDicer cells when compared with shNC cells following cisplatin treatment in the A2780 and CAOV3 cell lines. Western blot analyses also demonstrated that P21 protein expression was not affected by Dicer downregulation in A2780 and CAOV3 cells, while it was decreased following cisplatin treatment. The results were normalized to the expression of the reference protein β-actin. shRNA, short hairpin RNA; NC, negative control.

    Article Snippet: Membranes were incubated with mouse anti-Dicer (1:500 dilution; Abcam, Cambridge, MA, USA), rabbit anti-P53 (1:1,000 dilution; Cell Signaling Technology, Inc., Danvers, MA, USA), rabbit anti-P63 (1:500 dilution; Abcam), rabbit anti-P73 (1:500 dilution; Abcam), rabbit anti-caspase-9 (1:1,000 dilution; Cell Signaling Technology, Inc.), rabbit anti-caspase-3 (1:2,000 dilution; Cell Signaling Technology, Inc.), rabbit anti-P21 (1:1,000 dilution; Cell Signaling Technology, Inc.) and mouse anti-β-actin (1:5,000 dilution; Wuhan Boster Biological Technology, Ltd., Wuhan, China). β-Actin was used as a loading control.

    Techniques: Expressing, Western Blot, shRNA, Negative Control