etoposide Millipore Search Results


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  • 99
    Millipore etoposide
    Etoposide, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 6147 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 6147 article reviews
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    88
    Millipore etoposide vp 16
    Silencing of SNAI2 phenocopies the effects of miR-203 re-expression on sensitization of U251AR cells to anticancer drugs and reversion of EMT (A) a. U251AR/shNC cells; b. U251AR/shSNAI2 cells. Light microscopy, 100× (a, b); Fluorescent microscopy, 100× (a, b). shSNAI2 and negative vector (shNC) were transfected into U251AR cells. At 48 h after transfection, fluorescent microscopy showed emission green fluorescence. (B) qRT-PCR validate the downregulation of SNAI2 after shRNA knockdown in U251AR cells. (C) Immunofluorescence analysis of the endogenous SNAI2 protein (red, left panels) in U251AR cells transfected with shSNAI2 or negative vector. Nuclei are stained in blue with DAPI. Scale bar, 20 μm. (D) The sensitivities of U251AR and U251AR/shSNAI2 to different concentrations of TMZ, imatinib and <t>VP-16.</t> (E) Morphology of U251AR cells transfected with negative vector or shSNAI2 vector. Scale bar, 100 μm. (F) SNAI2 knockdown reduces the invasion capacity of U251AR cells. Scale bar, 200 μm. (G) U251AR cell monolayer was transfected as indicated and scratched, then the migration of the cells towards the wound was visualised. Images were taken at various time points and Image J was used to determine the migration distance. (H) Western blotting show that silencing of SNAI2 can modulate the expression of EMT markers. VP-16, <t>etoposide;</t> TMZ, temozolomide. Data are presented as mean±s.d. of three independent experiments. * P
    Etoposide Vp 16, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 305 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Millipore anticancer drug etoposide vp 16
    Silencing of SNAI2 phenocopies the effects of miR-203 re-expression on sensitization of U251AR cells to anticancer drugs and reversion of EMT (A) a. U251AR/shNC cells; b. U251AR/shSNAI2 cells. Light microscopy, 100× (a, b); Fluorescent microscopy, 100× (a, b). shSNAI2 and negative vector (shNC) were transfected into U251AR cells. At 48 h after transfection, fluorescent microscopy showed emission green fluorescence. (B) qRT-PCR validate the downregulation of SNAI2 after shRNA knockdown in U251AR cells. (C) Immunofluorescence analysis of the endogenous SNAI2 protein (red, left panels) in U251AR cells transfected with shSNAI2 or negative vector. Nuclei are stained in blue with DAPI. Scale bar, 20 μm. (D) The sensitivities of U251AR and U251AR/shSNAI2 to different concentrations of TMZ, imatinib and <t>VP-16.</t> (E) Morphology of U251AR cells transfected with negative vector or shSNAI2 vector. Scale bar, 100 μm. (F) SNAI2 knockdown reduces the invasion capacity of U251AR cells. Scale bar, 200 μm. (G) U251AR cell monolayer was transfected as indicated and scratched, then the migration of the cells towards the wound was visualised. Images were taken at various time points and Image J was used to determine the migration distance. (H) Western blotting show that silencing of SNAI2 can modulate the expression of EMT markers. VP-16, <t>etoposide;</t> TMZ, temozolomide. Data are presented as mean±s.d. of three independent experiments. * P
    Anticancer Drug Etoposide Vp 16, supplied by Millipore, used in various techniques. Bioz Stars score: 80/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Millipore etoposide eto
    Silencing of SNAI2 phenocopies the effects of miR-203 re-expression on sensitization of U251AR cells to anticancer drugs and reversion of EMT (A) a. U251AR/shNC cells; b. U251AR/shSNAI2 cells. Light microscopy, 100× (a, b); Fluorescent microscopy, 100× (a, b). shSNAI2 and negative vector (shNC) were transfected into U251AR cells. At 48 h after transfection, fluorescent microscopy showed emission green fluorescence. (B) qRT-PCR validate the downregulation of SNAI2 after shRNA knockdown in U251AR cells. (C) Immunofluorescence analysis of the endogenous SNAI2 protein (red, left panels) in U251AR cells transfected with shSNAI2 or negative vector. Nuclei are stained in blue with DAPI. Scale bar, 20 μm. (D) The sensitivities of U251AR and U251AR/shSNAI2 to different concentrations of TMZ, imatinib and <t>VP-16.</t> (E) Morphology of U251AR cells transfected with negative vector or shSNAI2 vector. Scale bar, 100 μm. (F) SNAI2 knockdown reduces the invasion capacity of U251AR cells. Scale bar, 200 μm. (G) U251AR cell monolayer was transfected as indicated and scratched, then the migration of the cells towards the wound was visualised. Images were taken at various time points and Image J was used to determine the migration distance. (H) Western blotting show that silencing of SNAI2 can modulate the expression of EMT markers. VP-16, <t>etoposide;</t> TMZ, temozolomide. Data are presented as mean±s.d. of three independent experiments. * P
    Etoposide Eto, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Millipore etoposide etp
    Silencing of SNAI2 phenocopies the effects of miR-203 re-expression on sensitization of U251AR cells to anticancer drugs and reversion of EMT (A) a. U251AR/shNC cells; b. U251AR/shSNAI2 cells. Light microscopy, 100× (a, b); Fluorescent microscopy, 100× (a, b). shSNAI2 and negative vector (shNC) were transfected into U251AR cells. At 48 h after transfection, fluorescent microscopy showed emission green fluorescence. (B) qRT-PCR validate the downregulation of SNAI2 after shRNA knockdown in U251AR cells. (C) Immunofluorescence analysis of the endogenous SNAI2 protein (red, left panels) in U251AR cells transfected with shSNAI2 or negative vector. Nuclei are stained in blue with DAPI. Scale bar, 20 μm. (D) The sensitivities of U251AR and U251AR/shSNAI2 to different concentrations of TMZ, imatinib and <t>VP-16.</t> (E) Morphology of U251AR cells transfected with negative vector or shSNAI2 vector. Scale bar, 100 μm. (F) SNAI2 knockdown reduces the invasion capacity of U251AR cells. Scale bar, 200 μm. (G) U251AR cell monolayer was transfected as indicated and scratched, then the migration of the cells towards the wound was visualised. Images were taken at various time points and Image J was used to determine the migration distance. (H) Western blotting show that silencing of SNAI2 can modulate the expression of EMT markers. VP-16, <t>etoposide;</t> TMZ, temozolomide. Data are presented as mean±s.d. of three independent experiments. * P
    Etoposide Etp, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Millipore m etoposide
    Silencing of SNAI2 phenocopies the effects of miR-203 re-expression on sensitization of U251AR cells to anticancer drugs and reversion of EMT (A) a. U251AR/shNC cells; b. U251AR/shSNAI2 cells. Light microscopy, 100× (a, b); Fluorescent microscopy, 100× (a, b). shSNAI2 and negative vector (shNC) were transfected into U251AR cells. At 48 h after transfection, fluorescent microscopy showed emission green fluorescence. (B) qRT-PCR validate the downregulation of SNAI2 after shRNA knockdown in U251AR cells. (C) Immunofluorescence analysis of the endogenous SNAI2 protein (red, left panels) in U251AR cells transfected with shSNAI2 or negative vector. Nuclei are stained in blue with DAPI. Scale bar, 20 μm. (D) The sensitivities of U251AR and U251AR/shSNAI2 to different concentrations of TMZ, imatinib and <t>VP-16.</t> (E) Morphology of U251AR cells transfected with negative vector or shSNAI2 vector. Scale bar, 100 μm. (F) SNAI2 knockdown reduces the invasion capacity of U251AR cells. Scale bar, 200 μm. (G) U251AR cell monolayer was transfected as indicated and scratched, then the migration of the cells towards the wound was visualised. Images were taken at various time points and Image J was used to determine the migration distance. (H) Western blotting show that silencing of SNAI2 can modulate the expression of EMT markers. VP-16, <t>etoposide;</t> TMZ, temozolomide. Data are presented as mean±s.d. of three independent experiments. * P
    M Etoposide, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 49 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Millipore 100um etoposide
    Silencing of SNAI2 phenocopies the effects of miR-203 re-expression on sensitization of U251AR cells to anticancer drugs and reversion of EMT (A) a. U251AR/shNC cells; b. U251AR/shSNAI2 cells. Light microscopy, 100× (a, b); Fluorescent microscopy, 100× (a, b). shSNAI2 and negative vector (shNC) were transfected into U251AR cells. At 48 h after transfection, fluorescent microscopy showed emission green fluorescence. (B) qRT-PCR validate the downregulation of SNAI2 after shRNA knockdown in U251AR cells. (C) Immunofluorescence analysis of the endogenous SNAI2 protein (red, left panels) in U251AR cells transfected with shSNAI2 or negative vector. Nuclei are stained in blue with DAPI. Scale bar, 20 μm. (D) The sensitivities of U251AR and U251AR/shSNAI2 to different concentrations of TMZ, imatinib and <t>VP-16.</t> (E) Morphology of U251AR cells transfected with negative vector or shSNAI2 vector. Scale bar, 100 μm. (F) SNAI2 knockdown reduces the invasion capacity of U251AR cells. Scale bar, 200 μm. (G) U251AR cell monolayer was transfected as indicated and scratched, then the migration of the cells towards the wound was visualised. Images were taken at various time points and Image J was used to determine the migration distance. (H) Western blotting show that silencing of SNAI2 can modulate the expression of EMT markers. VP-16, <t>etoposide;</t> TMZ, temozolomide. Data are presented as mean±s.d. of three independent experiments. * P
    100um Etoposide, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore etoposide vp16
    miR-3196 inhibits <t>VP-16</t> induced apoptosis by downregulating PUMA A. A549 cells were transfected with miR-3196 mimics (miR-3196) and/or combined with pcDNA3-PUMA (PUMA) for 24 h, followed by treatment with <t>VP16</t> (100 μM) for 48 h. The apoptotic cells were detected by FCM (left panel). Error bars denote the mean ± SD (right panel). B. H1650 cells were transfected and treated as in (A). The apoptotic cells were detected by FCM (left panel). Error bars denote the mean ± SD (right panel). * P
    Etoposide Vp16, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Millipore etoposide et
    miR-3196 inhibits <t>VP-16</t> induced apoptosis by downregulating PUMA A. A549 cells were transfected with miR-3196 mimics (miR-3196) and/or combined with pcDNA3-PUMA (PUMA) for 24 h, followed by treatment with <t>VP16</t> (100 μM) for 48 h. The apoptotic cells were detected by FCM (left panel). Error bars denote the mean ± SD (right panel). B. H1650 cells were transfected and treated as in (A). The apoptotic cells were detected by FCM (left panel). Error bars denote the mean ± SD (right panel). * P
    Etoposide Et, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    82
    Millipore etoposide control
    miR-3196 inhibits <t>VP-16</t> induced apoptosis by downregulating PUMA A. A549 cells were transfected with miR-3196 mimics (miR-3196) and/or combined with pcDNA3-PUMA (PUMA) for 24 h, followed by treatment with <t>VP16</t> (100 μM) for 48 h. The apoptotic cells were detected by FCM (left panel). Error bars denote the mean ± SD (right panel). B. H1650 cells were transfected and treated as in (A). The apoptotic cells were detected by FCM (left panel). Error bars denote the mean ± SD (right panel). * P
    Etoposide Control, supplied by Millipore, used in various techniques. Bioz Stars score: 82/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Millipore 10um etoposide
    miR-3196 inhibits <t>VP-16</t> induced apoptosis by downregulating PUMA A. A549 cells were transfected with miR-3196 mimics (miR-3196) and/or combined with pcDNA3-PUMA (PUMA) for 24 h, followed by treatment with <t>VP16</t> (100 μM) for 48 h. The apoptotic cells were detected by FCM (left panel). Error bars denote the mean ± SD (right panel). B. H1650 cells were transfected and treated as in (A). The apoptotic cells were detected by FCM (left panel). Error bars denote the mean ± SD (right panel). * P
    10um Etoposide, supplied by Millipore, used in various techniques. Bioz Stars score: 80/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Millipore rabbit anti vp 16
    GDNF and <t>VP-16</t> expression patterns . The GDNF expression pattern of the final ON group ( a , b ) was similar to that previously observed where the GDNF transgene (red) is observed distally from that of the transduced cells and VP-16 (green). ( b ) Increased magnification of area outlined in a . VP-16 expression in the OFF group ( c , d ) was similar to that seen in the ON group; however, no GDNF protein expression was observed. GDNF protein expression in the CBA-GDNF group ( e , f ) extended in a diffuse pattern into the VTA and the DMN indicating that GDNF protein was distributed away from the transduced cells in contrast to the pattern of expression observed from an intracellular transgene such as the SN-GFP group ( g , h ). As expected, no immunoreactivity was observed for VP-16 in either the CBA-GDNF group ( e , f , green) or the SN-GFP group ( g , h , red). ( f and h ) Higher magnifications of areas outlined in e and g , respectively. Bar: a , c , e , g = 1.0 mm, bar: b , d , f , h = 0.1 mm. CBA, chicken β-actin; DMN, deep mesencephalic nucleus; GDNF, glial cell line-derived neurotrophic factor; SNc, substantia nigra pars compacta; SNr, substantia nigra pars reticulate; VTA, ventral tegmental area.
    Rabbit Anti Vp 16, supplied by Millipore, used in various techniques. Bioz Stars score: 79/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Millipore etoposide atp agarose
    GDNF and <t>VP-16</t> expression patterns . The GDNF expression pattern of the final ON group ( a , b ) was similar to that previously observed where the GDNF transgene (red) is observed distally from that of the transduced cells and VP-16 (green). ( b ) Increased magnification of area outlined in a . VP-16 expression in the OFF group ( c , d ) was similar to that seen in the ON group; however, no GDNF protein expression was observed. GDNF protein expression in the CBA-GDNF group ( e , f ) extended in a diffuse pattern into the VTA and the DMN indicating that GDNF protein was distributed away from the transduced cells in contrast to the pattern of expression observed from an intracellular transgene such as the SN-GFP group ( g , h ). As expected, no immunoreactivity was observed for VP-16 in either the CBA-GDNF group ( e , f , green) or the SN-GFP group ( g , h , red). ( f and h ) Higher magnifications of areas outlined in e and g , respectively. Bar: a , c , e , g = 1.0 mm, bar: b , d , f , h = 0.1 mm. CBA, chicken β-actin; DMN, deep mesencephalic nucleus; GDNF, glial cell line-derived neurotrophic factor; SNc, substantia nigra pars compacta; SNr, substantia nigra pars reticulate; VTA, ventral tegmental area.
    Etoposide Atp Agarose, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Millipore daunorubicin analogue vp 16
    GDNF and <t>VP-16</t> expression patterns . The GDNF expression pattern of the final ON group ( a , b ) was similar to that previously observed where the GDNF transgene (red) is observed distally from that of the transduced cells and VP-16 (green). ( b ) Increased magnification of area outlined in a . VP-16 expression in the OFF group ( c , d ) was similar to that seen in the ON group; however, no GDNF protein expression was observed. GDNF protein expression in the CBA-GDNF group ( e , f ) extended in a diffuse pattern into the VTA and the DMN indicating that GDNF protein was distributed away from the transduced cells in contrast to the pattern of expression observed from an intracellular transgene such as the SN-GFP group ( g , h ). As expected, no immunoreactivity was observed for VP-16 in either the CBA-GDNF group ( e , f , green) or the SN-GFP group ( g , h , red). ( f and h ) Higher magnifications of areas outlined in e and g , respectively. Bar: a , c , e , g = 1.0 mm, bar: b , d , f , h = 0.1 mm. CBA, chicken β-actin; DMN, deep mesencephalic nucleus; GDNF, glial cell line-derived neurotrophic factor; SNc, substantia nigra pars compacta; SNr, substantia nigra pars reticulate; VTA, ventral tegmental area.
    Daunorubicin Analogue Vp 16, supplied by Millipore, used in various techniques. Bioz Stars score: 80/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Millipore bone marrow suppressant vp 16
    Effects of engraftment location, estrogen, and immune suppressant on tumorigenicity of breast cancer cells NOD/SCID mice were treated ± 17β-estradiol (E 2 ) and <t>etoposide</t> as indicated. Six days later, mice were surgically incised and injected with 500 cells from a breast cancer pleural effusion in 25% Matrigel. A: final tumor volume. Data represent mean ± SE. B: tumor incidence. C: latency of tumor formation following injection (* P
    Bone Marrow Suppressant Vp 16, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore topoisomerase ii inhibitor etoposide
    Effects of engraftment location, estrogen, and immune suppressant on tumorigenicity of breast cancer cells NOD/SCID mice were treated ± 17β-estradiol (E 2 ) and <t>etoposide</t> as indicated. Six days later, mice were surgically incised and injected with 500 cells from a breast cancer pleural effusion in 25% Matrigel. A: final tumor volume. Data represent mean ± SE. B: tumor incidence. C: latency of tumor formation following injection (* P
    Topoisomerase Ii Inhibitor Etoposide, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 65 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore drug resistance against etoposide
    Effects of engraftment location, estrogen, and immune suppressant on tumorigenicity of breast cancer cells NOD/SCID mice were treated ± 17β-estradiol (E 2 ) and <t>etoposide</t> as indicated. Six days later, mice were surgically incised and injected with 500 cells from a breast cancer pleural effusion in 25% Matrigel. A: final tumor volume. Data represent mean ± SE. B: tumor incidence. C: latency of tumor formation following injection (* P
    Drug Resistance Against Etoposide, supplied by Millipore, used in various techniques. Bioz Stars score: 79/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Millipore etoposide vp16 etp
    The combined use of anti-CD33 mAb and <t>etoposide</t> displays a synergistic effect in binding of Annexin V. The surface binding of Annexin V to AML cells was measured after 20 h of culture with GM-CSF in the presence of anti-CD33 mAb (μg/ml), <t>ETP</t> (68 μM/ml), or ETP (34 μM/ml) used either alone or in combination. The experiment shown is representative of four independent experiments. Samples were stained with Annexin V FITC-conjugated mAb. Analysis was performed on PI-negative, viable cells.
    Etoposide Vp16 Etp, supplied by Millipore, used in various techniques. Bioz Stars score: 77/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Millipore topoisomerase 2 poisons etoposide
    The combined use of anti-CD33 mAb and <t>etoposide</t> displays a synergistic effect in binding of Annexin V. The surface binding of Annexin V to AML cells was measured after 20 h of culture with GM-CSF in the presence of anti-CD33 mAb (μg/ml), <t>ETP</t> (68 μM/ml), or ETP (34 μM/ml) used either alone or in combination. The experiment shown is representative of four independent experiments. Samples were stained with Annexin V FITC-conjugated mAb. Analysis was performed on PI-negative, viable cells.
    Topoisomerase 2 Poisons Etoposide, supplied by Millipore, used in various techniques. Bioz Stars score: 77/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Millipore apoptosis stock etoposide
    The combined use of anti-CD33 mAb and <t>etoposide</t> displays a synergistic effect in binding of Annexin V. The surface binding of Annexin V to AML cells was measured after 20 h of culture with GM-CSF in the presence of anti-CD33 mAb (μg/ml), <t>ETP</t> (68 μM/ml), or ETP (34 μM/ml) used either alone or in combination. The experiment shown is representative of four independent experiments. Samples were stained with Annexin V FITC-conjugated mAb. Analysis was performed on PI-negative, viable cells.
    Apoptosis Stock Etoposide, supplied by Millipore, used in various techniques. Bioz Stars score: 77/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    81
    Millipore topo ii inhibitor etoposide
    MMS22L foci form during S phase of the cell cycle and colocalize with HR proteins Immunofluorescence images of MMS22L foci formation at <t>etoposide</t> (ETP)‐induced DSBs during S, M, and G1 phases of the cell cycle in HeLa cells. Scale bar: 5 μm. Quantification of a representative experiment from (A) shows number of MMS22L foci per nucleus during indicated cell cycle phases ( n = 2; n nuclei ≥ 60). Immunofluorescence images of CPT‐induced MMS22L, RPA2, BRCA2, and RAD51 foci in U2OS cells. Scale bar: 5 μm. Quantification of a representative experiment ( n = 3) from (C) shows percentage of γH2AX, RPA2, BRCA2, and RAD51 foci colocalizing with MMS22L foci ( n nuclei = 30; nuclei with ≥ 25 MMS22L foci were analyzed). Data information: Boxplots in (B and D) represent distributions per nucleus; boxes indicate the 25–75 percentile and whiskers the 10–90 percentile. Horizontal lines mark the medians. Statistical analysis: Mann–Whitney U ‐test; *** P ≤ 0.0001; ns, not significant.
    Topo Ii Inhibitor Etoposide, supplied by Millipore, used in various techniques. Bioz Stars score: 81/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore clonogenic survival assay etoposide
    MMS22L foci form during S phase of the cell cycle and colocalize with HR proteins Immunofluorescence images of MMS22L foci formation at <t>etoposide</t> (ETP)‐induced DSBs during S, M, and G1 phases of the cell cycle in HeLa cells. Scale bar: 5 μm. Quantification of a representative experiment from (A) shows number of MMS22L foci per nucleus during indicated cell cycle phases ( n = 2; n nuclei ≥ 60). Immunofluorescence images of CPT‐induced MMS22L, RPA2, BRCA2, and RAD51 foci in U2OS cells. Scale bar: 5 μm. Quantification of a representative experiment ( n = 3) from (C) shows percentage of γH2AX, RPA2, BRCA2, and RAD51 foci colocalizing with MMS22L foci ( n nuclei = 30; nuclei with ≥ 25 MMS22L foci were analyzed). Data information: Boxplots in (B and D) represent distributions per nucleus; boxes indicate the 25–75 percentile and whiskers the 10–90 percentile. Horizontal lines mark the medians. Statistical analysis: Mann–Whitney U ‐test; *** P ≤ 0.0001; ns, not significant.
    Clonogenic Survival Assay Etoposide, supplied by Millipore, used in various techniques. Bioz Stars score: 79/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MMS22L foci form during S phase of the cell cycle and colocalize with HR proteins Immunofluorescence images of MMS22L foci formation at <t>etoposide</t> (ETP)‐induced DSBs during S, M, and G1 phases of the cell cycle in HeLa cells. Scale bar: 5 μm. Quantification of a representative experiment from (A) shows number of MMS22L foci per nucleus during indicated cell cycle phases ( n = 2; n nuclei ≥ 60). Immunofluorescence images of CPT‐induced MMS22L, RPA2, BRCA2, and RAD51 foci in U2OS cells. Scale bar: 5 μm. Quantification of a representative experiment ( n = 3) from (C) shows percentage of γH2AX, RPA2, BRCA2, and RAD51 foci colocalizing with MMS22L foci ( n nuclei = 30; nuclei with ≥ 25 MMS22L foci were analyzed). Data information: Boxplots in (B and D) represent distributions per nucleus; boxes indicate the 25–75 percentile and whiskers the 10–90 percentile. Horizontal lines mark the medians. Statistical analysis: Mann–Whitney U ‐test; *** P ≤ 0.0001; ns, not significant.
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    MMS22L foci form during S phase of the cell cycle and colocalize with HR proteins Immunofluorescence images of MMS22L foci formation at <t>etoposide</t> (ETP)‐induced DSBs during S, M, and G1 phases of the cell cycle in HeLa cells. Scale bar: 5 μm. Quantification of a representative experiment from (A) shows number of MMS22L foci per nucleus during indicated cell cycle phases ( n = 2; n nuclei ≥ 60). Immunofluorescence images of CPT‐induced MMS22L, RPA2, BRCA2, and RAD51 foci in U2OS cells. Scale bar: 5 μm. Quantification of a representative experiment ( n = 3) from (C) shows percentage of γH2AX, RPA2, BRCA2, and RAD51 foci colocalizing with MMS22L foci ( n nuclei = 30; nuclei with ≥ 25 MMS22L foci were analyzed). Data information: Boxplots in (B and D) represent distributions per nucleus; boxes indicate the 25–75 percentile and whiskers the 10–90 percentile. Horizontal lines mark the medians. Statistical analysis: Mann–Whitney U ‐test; *** P ≤ 0.0001; ns, not significant.
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    MMS22L foci form during S phase of the cell cycle and colocalize with HR proteins Immunofluorescence images of MMS22L foci formation at <t>etoposide</t> (ETP)‐induced DSBs during S, M, and G1 phases of the cell cycle in HeLa cells. Scale bar: 5 μm. Quantification of a representative experiment from (A) shows number of MMS22L foci per nucleus during indicated cell cycle phases ( n = 2; n nuclei ≥ 60). Immunofluorescence images of CPT‐induced MMS22L, RPA2, BRCA2, and RAD51 foci in U2OS cells. Scale bar: 5 μm. Quantification of a representative experiment ( n = 3) from (C) shows percentage of γH2AX, RPA2, BRCA2, and RAD51 foci colocalizing with MMS22L foci ( n nuclei = 30; nuclei with ≥ 25 MMS22L foci were analyzed). Data information: Boxplots in (B and D) represent distributions per nucleus; boxes indicate the 25–75 percentile and whiskers the 10–90 percentile. Horizontal lines mark the medians. Statistical analysis: Mann–Whitney U ‐test; *** P ≤ 0.0001; ns, not significant.
    Atm Atr Signaling Cascade Analysis Etoposide, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore etoposide 4 demethylepipodophyllotoxin 9 4 6 o ethylidene β d glucopyranoside
    MMS22L foci form during S phase of the cell cycle and colocalize with HR proteins Immunofluorescence images of MMS22L foci formation at <t>etoposide</t> (ETP)‐induced DSBs during S, M, and G1 phases of the cell cycle in HeLa cells. Scale bar: 5 μm. Quantification of a representative experiment from (A) shows number of MMS22L foci per nucleus during indicated cell cycle phases ( n = 2; n nuclei ≥ 60). Immunofluorescence images of CPT‐induced MMS22L, RPA2, BRCA2, and RAD51 foci in U2OS cells. Scale bar: 5 μm. Quantification of a representative experiment ( n = 3) from (C) shows percentage of γH2AX, RPA2, BRCA2, and RAD51 foci colocalizing with MMS22L foci ( n nuclei = 30; nuclei with ≥ 25 MMS22L foci were analyzed). Data information: Boxplots in (B and D) represent distributions per nucleus; boxes indicate the 25–75 percentile and whiskers the 10–90 percentile. Horizontal lines mark the medians. Statistical analysis: Mann–Whitney U ‐test; *** P ≤ 0.0001; ns, not significant.
    Etoposide 4 Demethylepipodophyllotoxin 9 4 6 O Ethylidene β D Glucopyranoside, supplied by Millipore, used in various techniques. Bioz Stars score: 77/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Flow cytometry analysis of the rates of apoptosis in Huh7 cells exposed to DIC or to IND . For apoptosis analysis, permeabilized Huh7 cells were labelled with propidium iodide (PI) and analysed by flow cytometry by a FACSCalibur flow cytometer (Becton Dickinson, North Ryde, NSW, Australia). DNA damaging agent <t>etopoxide</t> was used as positive control for apoptosis. Results are the mean of three experiments repeated three times.
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    Flow cytometry analysis of the rates of apoptosis in Huh7 cells exposed to DIC or to IND . For apoptosis analysis, permeabilized Huh7 cells were labelled with propidium iodide (PI) and analysed by flow cytometry by a FACSCalibur flow cytometer (Becton Dickinson, North Ryde, NSW, Australia). DNA damaging agent <t>etopoxide</t> was used as positive control for apoptosis. Results are the mean of three experiments repeated three times.
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    Precise quantification of individual treatment effects and monitoring of distinct clinical stages. a Imaging visualizes treatment-induced cell loss and regrowth; 10 5 ALL-50 cells/mouse were injected into 10 mice which received a single intraperitoneal dose of <t>Etoposid</t> (VP-16; 50 mg/kg) in week 6 after tumor cell injection, except the control mouse which was treated with PBS. Animals were imaged before treatment (pre-treatment) and 4 and 11 days after treatment; shown is one representative mouse; all mice are shown in Supplemental Figure 11; b, c Imaging visualizes different sensitivities of individual samples towards treatment; ALL-199 (1×10 4 cells/mouse) or ALL-4S (5×10 4 cells/mouse) were injected into 16 mice; mice were randomized in week 4 into one control (n = 4) and two experimental groups (n = 6 each). Control mice received buffer injection, while the other groups were treated once intraperitoneally with either Etoposid (VP-16; 50 mg/kg) or cyclophosphamide (Cyclo; 150 mg/kg) as indicated. Mice were imaged directly before and 4 days after treatment; shown are 3 representative mice of each treatment group before and 4 days after treatment ( b ); shown is the result of quantification of the images; each line represents a single mouse ( c ); * P
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    Precise quantification of individual treatment effects and monitoring of distinct clinical stages. a Imaging visualizes treatment-induced cell loss and regrowth; 10 5 ALL-50 cells/mouse were injected into 10 mice which received a single intraperitoneal dose of <t>Etoposid</t> (VP-16; 50 mg/kg) in week 6 after tumor cell injection, except the control mouse which was treated with PBS. Animals were imaged before treatment (pre-treatment) and 4 and 11 days after treatment; shown is one representative mouse; all mice are shown in Supplemental Figure 11; b, c Imaging visualizes different sensitivities of individual samples towards treatment; ALL-199 (1×10 4 cells/mouse) or ALL-4S (5×10 4 cells/mouse) were injected into 16 mice; mice were randomized in week 4 into one control (n = 4) and two experimental groups (n = 6 each). Control mice received buffer injection, while the other groups were treated once intraperitoneally with either Etoposid (VP-16; 50 mg/kg) or cyclophosphamide (Cyclo; 150 mg/kg) as indicated. Mice were imaged directly before and 4 days after treatment; shown are 3 representative mice of each treatment group before and 4 days after treatment ( b ); shown is the result of quantification of the images; each line represents a single mouse ( c ); * P
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    Image Search Results


    Silencing of SNAI2 phenocopies the effects of miR-203 re-expression on sensitization of U251AR cells to anticancer drugs and reversion of EMT (A) a. U251AR/shNC cells; b. U251AR/shSNAI2 cells. Light microscopy, 100× (a, b); Fluorescent microscopy, 100× (a, b). shSNAI2 and negative vector (shNC) were transfected into U251AR cells. At 48 h after transfection, fluorescent microscopy showed emission green fluorescence. (B) qRT-PCR validate the downregulation of SNAI2 after shRNA knockdown in U251AR cells. (C) Immunofluorescence analysis of the endogenous SNAI2 protein (red, left panels) in U251AR cells transfected with shSNAI2 or negative vector. Nuclei are stained in blue with DAPI. Scale bar, 20 μm. (D) The sensitivities of U251AR and U251AR/shSNAI2 to different concentrations of TMZ, imatinib and VP-16. (E) Morphology of U251AR cells transfected with negative vector or shSNAI2 vector. Scale bar, 100 μm. (F) SNAI2 knockdown reduces the invasion capacity of U251AR cells. Scale bar, 200 μm. (G) U251AR cell monolayer was transfected as indicated and scratched, then the migration of the cells towards the wound was visualised. Images were taken at various time points and Image J was used to determine the migration distance. (H) Western blotting show that silencing of SNAI2 can modulate the expression of EMT markers. VP-16, etoposide; TMZ, temozolomide. Data are presented as mean±s.d. of three independent experiments. * P

    Journal: Oncotarget

    Article Title: MiR-203 downregulation is responsible for chemoresistance in human glioblastoma by promoting epithelial-mesenchymal transition via SNAI2

    doi:

    Figure Lengend Snippet: Silencing of SNAI2 phenocopies the effects of miR-203 re-expression on sensitization of U251AR cells to anticancer drugs and reversion of EMT (A) a. U251AR/shNC cells; b. U251AR/shSNAI2 cells. Light microscopy, 100× (a, b); Fluorescent microscopy, 100× (a, b). shSNAI2 and negative vector (shNC) were transfected into U251AR cells. At 48 h after transfection, fluorescent microscopy showed emission green fluorescence. (B) qRT-PCR validate the downregulation of SNAI2 after shRNA knockdown in U251AR cells. (C) Immunofluorescence analysis of the endogenous SNAI2 protein (red, left panels) in U251AR cells transfected with shSNAI2 or negative vector. Nuclei are stained in blue with DAPI. Scale bar, 20 μm. (D) The sensitivities of U251AR and U251AR/shSNAI2 to different concentrations of TMZ, imatinib and VP-16. (E) Morphology of U251AR cells transfected with negative vector or shSNAI2 vector. Scale bar, 100 μm. (F) SNAI2 knockdown reduces the invasion capacity of U251AR cells. Scale bar, 200 μm. (G) U251AR cell monolayer was transfected as indicated and scratched, then the migration of the cells towards the wound was visualised. Images were taken at various time points and Image J was used to determine the migration distance. (H) Western blotting show that silencing of SNAI2 can modulate the expression of EMT markers. VP-16, etoposide; TMZ, temozolomide. Data are presented as mean±s.d. of three independent experiments. * P

    Article Snippet: After 24 h, the cells were treated with different concentrations of imatinib (Novartis, Basel, Switzerland), etoposide (VP-16) (Sigma Chemical Co., St. Louis, MO) and temozolomide (TMZ) (Sigma Chemical Co., St. Louis, MO), each at four concentrations ranging from 50 to 200 μg/ml for 48 h. The range of drug concentrations were based on earlier studies and aimed at obtaining IC50 values both for highly sensitive and resistant cases.

    Techniques: Expressing, Light Microscopy, Microscopy, Plasmid Preparation, Transfection, Fluorescence, Quantitative RT-PCR, shRNA, Immunofluorescence, Staining, Migration, Western Blot

    Re-expression of miR-203 in U251AR and U87AR cells sensitizes cells to anticancer drugs and reverses EMT while knockdown of miR-203 promotes resistance to anticancer drugs in U251 and U87 cells (A) qRT-PCR data validation of the downregulation of miR-203 in imatinib-resistant GBM cells compared with their parental cells, normalized to U6RNA, which was obtained from miRNA microarrays. (B, C) The sensitivities of U251AR and U87AR cells to imatinib, VP-16 and TMZ after transfected with miR-203 or miRNAs control. (D, E) Transfection with anti-miR-203 promotes resistance to imatinib, VP-16 and TMZ in U251 and U87 cells. (F) Morphology of U251AR and U87AR cells transfected with miRNA control or miR-203. Scale bar, 100 μm. (G) Western blotting show that re-expression of miR-203 modulates the expression of EMT markers. (H, I) U251AR and U87AR cells were transfected with miR-203 or anti-miR-203, and then collected for transwell invasion assay or wound healing assay. Shown were pictures of representative fields for each experiment. Scale bar, 200 μm. Data were expressed as mean±s.d. from three independent experiments. VP-16, etoposide; TMZ, temozolomide. * P

    Journal: Oncotarget

    Article Title: MiR-203 downregulation is responsible for chemoresistance in human glioblastoma by promoting epithelial-mesenchymal transition via SNAI2

    doi:

    Figure Lengend Snippet: Re-expression of miR-203 in U251AR and U87AR cells sensitizes cells to anticancer drugs and reverses EMT while knockdown of miR-203 promotes resistance to anticancer drugs in U251 and U87 cells (A) qRT-PCR data validation of the downregulation of miR-203 in imatinib-resistant GBM cells compared with their parental cells, normalized to U6RNA, which was obtained from miRNA microarrays. (B, C) The sensitivities of U251AR and U87AR cells to imatinib, VP-16 and TMZ after transfected with miR-203 or miRNAs control. (D, E) Transfection with anti-miR-203 promotes resistance to imatinib, VP-16 and TMZ in U251 and U87 cells. (F) Morphology of U251AR and U87AR cells transfected with miRNA control or miR-203. Scale bar, 100 μm. (G) Western blotting show that re-expression of miR-203 modulates the expression of EMT markers. (H, I) U251AR and U87AR cells were transfected with miR-203 or anti-miR-203, and then collected for transwell invasion assay or wound healing assay. Shown were pictures of representative fields for each experiment. Scale bar, 200 μm. Data were expressed as mean±s.d. from three independent experiments. VP-16, etoposide; TMZ, temozolomide. * P

    Article Snippet: After 24 h, the cells were treated with different concentrations of imatinib (Novartis, Basel, Switzerland), etoposide (VP-16) (Sigma Chemical Co., St. Louis, MO) and temozolomide (TMZ) (Sigma Chemical Co., St. Louis, MO), each at four concentrations ranging from 50 to 200 μg/ml for 48 h. The range of drug concentrations were based on earlier studies and aimed at obtaining IC50 values both for highly sensitive and resistant cases.

    Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot, Transwell Invasion Assay, Wound Healing Assay

    SNAI2 contributes to chemoresistance and EMT in GBM cells (A) Overexpression of SNAI2 promotes resistance to imatinib, VP-16 and TMZ. (B) Morphology of U87 cells transfected with pcDNA3.1-mock or pcDNA3.1-SNAI2. Scale bar, 100 μm. (C) Invasion of U87 cells after pcDNA3.1-SNAI2 transfection. Scale bar, 200 μm. (D) Protein expression of EMT markers in U87 cells transfected with pcDNA3.1-mock or pcDNA3.1-SNAI2, determined by western blotting. (E) The sensitivities to imatinib, VP-16 and TMZ were measured after cells transfected with indicated constructs and miR-203 in U251AR. (F) Invasion assay of U251AR cells expressing indicated vectors and miR-203. (G) qRT-PCR for EMT markers in U251AR cells expressing indicated constructs and miR-203. * P

    Journal: Oncotarget

    Article Title: MiR-203 downregulation is responsible for chemoresistance in human glioblastoma by promoting epithelial-mesenchymal transition via SNAI2

    doi:

    Figure Lengend Snippet: SNAI2 contributes to chemoresistance and EMT in GBM cells (A) Overexpression of SNAI2 promotes resistance to imatinib, VP-16 and TMZ. (B) Morphology of U87 cells transfected with pcDNA3.1-mock or pcDNA3.1-SNAI2. Scale bar, 100 μm. (C) Invasion of U87 cells after pcDNA3.1-SNAI2 transfection. Scale bar, 200 μm. (D) Protein expression of EMT markers in U87 cells transfected with pcDNA3.1-mock or pcDNA3.1-SNAI2, determined by western blotting. (E) The sensitivities to imatinib, VP-16 and TMZ were measured after cells transfected with indicated constructs and miR-203 in U251AR. (F) Invasion assay of U251AR cells expressing indicated vectors and miR-203. (G) qRT-PCR for EMT markers in U251AR cells expressing indicated constructs and miR-203. * P

    Article Snippet: After 24 h, the cells were treated with different concentrations of imatinib (Novartis, Basel, Switzerland), etoposide (VP-16) (Sigma Chemical Co., St. Louis, MO) and temozolomide (TMZ) (Sigma Chemical Co., St. Louis, MO), each at four concentrations ranging from 50 to 200 μg/ml for 48 h. The range of drug concentrations were based on earlier studies and aimed at obtaining IC50 values both for highly sensitive and resistant cases.

    Techniques: Over Expression, Transfection, Expressing, Western Blot, Construct, Invasion Assay, Quantitative RT-PCR

    Zap70 and Syk loss is a consequence of DNA damage, but not of caspase activation (A) Representative flow cytometry plots showing the proportions of isolated NK cells positive for anti-cleaved Caspase 3 antibody (top) or NK cells that are Zap70 low (bottom). NK cells from a single donor, either treated with DMSO (left column, control) or with 0.25mg/mL VP-16 (right column) in DMSO, for 24 hours. Data are representative of 17 donors. (B) Percent of either Syk low , Zap70 low or cleaved Caspase 3 + in isolated NK cells from 9 donors following 24 hour treatment with 0.25mg/mL VP-16 etoposide, or DMSO control. (C) Percent of cleaved Caspase 3 + NK cells from 6 donors following 24 hour treatment with: DMSO control, 15uM Z-VAD-FMK caspase inhibitor alone, 0.25mg/mL VP-16 etoposide, or Z-VAD-FMK inhibitor and VP16 together. All cultures contained 500U/ml IL-2. Shown is the result of Tukey’s post test of paired ANOVA. (D) Percent of Zap70 low NK cells following 24 hour treatment, as in C, for seven donors. All panels show data that are representative of at least 3 independent experiments using NK cells from different donors. Statistical comparisons shown in all panels are the result of Tukey post-tests from paired 1-way or 2-way ANOVA, as appropriate. SEM are shown in all panels. **** = p

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Human Natural Killer Cells Downregulate Zap70 and Syk in Response to Prolonged Activation or DNA Damage

    doi: 10.4049/jimmunol.1700542

    Figure Lengend Snippet: Zap70 and Syk loss is a consequence of DNA damage, but not of caspase activation (A) Representative flow cytometry plots showing the proportions of isolated NK cells positive for anti-cleaved Caspase 3 antibody (top) or NK cells that are Zap70 low (bottom). NK cells from a single donor, either treated with DMSO (left column, control) or with 0.25mg/mL VP-16 (right column) in DMSO, for 24 hours. Data are representative of 17 donors. (B) Percent of either Syk low , Zap70 low or cleaved Caspase 3 + in isolated NK cells from 9 donors following 24 hour treatment with 0.25mg/mL VP-16 etoposide, or DMSO control. (C) Percent of cleaved Caspase 3 + NK cells from 6 donors following 24 hour treatment with: DMSO control, 15uM Z-VAD-FMK caspase inhibitor alone, 0.25mg/mL VP-16 etoposide, or Z-VAD-FMK inhibitor and VP16 together. All cultures contained 500U/ml IL-2. Shown is the result of Tukey’s post test of paired ANOVA. (D) Percent of Zap70 low NK cells following 24 hour treatment, as in C, for seven donors. All panels show data that are representative of at least 3 independent experiments using NK cells from different donors. Statistical comparisons shown in all panels are the result of Tukey post-tests from paired 1-way or 2-way ANOVA, as appropriate. SEM are shown in all panels. **** = p

    Article Snippet: In experiments to study DNA damage and apoptosis in NK cells, VP-16 etoposide (Sigma Aldrich) and/or Z-VAD FMK (BD Biosciences) were included in the culture medium at concentrations of 0.25mg/ml and 15μM, respectively.

    Techniques: Activation Assay, Flow Cytometry, Cytometry, Isolation

    The normalized, relative expression of CDKN1A /p21 after DMSO or 8 hr of etoposide treatment (100 μM final) in a pilot RT-qPCR screen and in the experimental siRNA screen (A). Average (B) CDKN1A /p21, (C) BBC3 /puma, and (D) TP53 /p53 expression values under DMSO and etoposide conditions across each experimental RT-qPCR plate for nontargeting, TP53 , and MDM2 siRNA. Results from T-tests of pairwise comparisons can be found in Table 1 . DMSO, dimethyl sulfoxide; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; siRNA, small interfering RNA.

    Journal: G3: Genes|Genomes|Genetics

    Article Title: A Chromatin-Focused siRNA Screen for Regulators of p53-Dependent Transcription

    doi: 10.1534/g3.116.031534

    Figure Lengend Snippet: The normalized, relative expression of CDKN1A /p21 after DMSO or 8 hr of etoposide treatment (100 μM final) in a pilot RT-qPCR screen and in the experimental siRNA screen (A). Average (B) CDKN1A /p21, (C) BBC3 /puma, and (D) TP53 /p53 expression values under DMSO and etoposide conditions across each experimental RT-qPCR plate for nontargeting, TP53 , and MDM2 siRNA. Results from T-tests of pairwise comparisons can be found in Table 1 . DMSO, dimethyl sulfoxide; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; siRNA, small interfering RNA.

    Article Snippet: Each siRNA was delivered to 11,000 cells via reverse transfection using RNAiMax (Life Technologies) in a 96-well plate to a final concentration of 10 μM, media was changed after 24 hr, and cells were incubated for an additional 48 hr before addition of either DMSO or 100 μM (final) etoposide (Sigma-Aldrich) for 8 hr.

    Techniques: Expressing, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Small Interfering RNA

    TCP80 and RHA interact in vivo and cooperatively stimulate p53 expression in MCF-7 cells. (a) TCP80 has increased binding with RHA following DNA damage in MCF-7 cells. Subconfluent MCF-7 cells were treated with or without 10 μ M etoposide for 2 hours and then lysed with TGN buffer [ 8 ]. RHA was immunoprecipitated from the cell lysate as described in experimental procedures. The precipitated beads were then washed three times with TGN lysis buffer and SDS sample loading buffer was added. The samples were subjected to SDS-PAGE. An immunoblotting experiment was then performed to detect the TCP80 protein. The results presented are representative of three individual experiments. (b) Levels of TCP80 and RHA protein do not change following exposure to DNA damage in MCF-7 cells. MCF-7 cells were treated with 10 μ M etoposide for 2 hours and then lysed with TGN lysis buffer. The samples were subjected to SDS-PAGE. TCP80, RHA, and β -actin were detected by their respective antibodies. The results presented in (a) and (b) are representative of three individual experiments. (c) Overexpression of TCP80 and RHA leads to increased p53 expression in H1299 cells transfected with the pC53-SN3 vector. H1299 lung carcinoma cells (p53-null) were cotransfected with the p53 expression vector pC53-SN3 along with the empty pCDNA 3.1 vector, the TCP80 expression vector, or the TCP80 plus RHA expression vector. Twenty-four hours after transfection, the cells were treated with or without etoposide for 2 hours. Cells were then lysed, and equal amounts of protein were subjected to SDS-PAGE and transferred to a nitrocellulose membrane. The p53 protein and β -actin were then detected by their respective antibodies. (d) Statistical analysis of the expression levels of p53 (p53/ β -actin) between individual groups as shown in (c) was performed using one-way ANOVA with a Newman–Keul post hoc test from 4 sets of experimental results. Significance was assumed at ∗ P

    Journal: BioMed Research International

    Article Title: Translational Control Protein 80 Stimulates IRES-Mediated Translation of p53 mRNA in Response to DNA Damage

    doi: 10.1155/2015/708158

    Figure Lengend Snippet: TCP80 and RHA interact in vivo and cooperatively stimulate p53 expression in MCF-7 cells. (a) TCP80 has increased binding with RHA following DNA damage in MCF-7 cells. Subconfluent MCF-7 cells were treated with or without 10 μ M etoposide for 2 hours and then lysed with TGN buffer [ 8 ]. RHA was immunoprecipitated from the cell lysate as described in experimental procedures. The precipitated beads were then washed three times with TGN lysis buffer and SDS sample loading buffer was added. The samples were subjected to SDS-PAGE. An immunoblotting experiment was then performed to detect the TCP80 protein. The results presented are representative of three individual experiments. (b) Levels of TCP80 and RHA protein do not change following exposure to DNA damage in MCF-7 cells. MCF-7 cells were treated with 10 μ M etoposide for 2 hours and then lysed with TGN lysis buffer. The samples were subjected to SDS-PAGE. TCP80, RHA, and β -actin were detected by their respective antibodies. The results presented in (a) and (b) are representative of three individual experiments. (c) Overexpression of TCP80 and RHA leads to increased p53 expression in H1299 cells transfected with the pC53-SN3 vector. H1299 lung carcinoma cells (p53-null) were cotransfected with the p53 expression vector pC53-SN3 along with the empty pCDNA 3.1 vector, the TCP80 expression vector, or the TCP80 plus RHA expression vector. Twenty-four hours after transfection, the cells were treated with or without etoposide for 2 hours. Cells were then lysed, and equal amounts of protein were subjected to SDS-PAGE and transferred to a nitrocellulose membrane. The p53 protein and β -actin were then detected by their respective antibodies. (d) Statistical analysis of the expression levels of p53 (p53/ β -actin) between individual groups as shown in (c) was performed using one-way ANOVA with a Newman–Keul post hoc test from 4 sets of experimental results. Significance was assumed at ∗ P

    Article Snippet: Materials Etoposide was from Calbiochem.

    Techniques: In Vivo, Expressing, Binding Assay, Immunoprecipitation, Lysis, SDS Page, Over Expression, Transfection, Plasmid Preparation

    (a) MCF-7/shTCP80 cells express lower levels of TCP80 and RHA as compared to MCF-7 cells. MCF-7 and MCF-7/shTCP80 cells were grown to subconfluency. Cells were then lysed and equal amounts of protein were subjected to SDS-PAGE and western blotting. TCP80, RHA, and β -actin were detected by immunoblotting. (b) MCF-7/shTCP80 cells exhibit reduced induction of p53 and its downstream target PUMA following DNA damage. MCF-7 and MCF-7/shTCP80 cells were grown to subconfluency. Cells were then treated with 10 μ M etoposide for 2 hours. After the treatment, cells were lysed and equal amounts of protein were subjected to SDS-PAGE and transferred to PVDF membranes. p53, PUMA, and β -actin proteins were detected with their respective antibodies. (c) Statistical analysis of the expression levels of p53 (p53/ β -actin) between individual groups as seen in (b) was carried out using one-way ANOVA with a Newman–Keul post hoc test from 3 sets of experimental results. Significance was assumed at ∗ P

    Journal: BioMed Research International

    Article Title: Translational Control Protein 80 Stimulates IRES-Mediated Translation of p53 mRNA in Response to DNA Damage

    doi: 10.1155/2015/708158

    Figure Lengend Snippet: (a) MCF-7/shTCP80 cells express lower levels of TCP80 and RHA as compared to MCF-7 cells. MCF-7 and MCF-7/shTCP80 cells were grown to subconfluency. Cells were then lysed and equal amounts of protein were subjected to SDS-PAGE and western blotting. TCP80, RHA, and β -actin were detected by immunoblotting. (b) MCF-7/shTCP80 cells exhibit reduced induction of p53 and its downstream target PUMA following DNA damage. MCF-7 and MCF-7/shTCP80 cells were grown to subconfluency. Cells were then treated with 10 μ M etoposide for 2 hours. After the treatment, cells were lysed and equal amounts of protein were subjected to SDS-PAGE and transferred to PVDF membranes. p53, PUMA, and β -actin proteins were detected with their respective antibodies. (c) Statistical analysis of the expression levels of p53 (p53/ β -actin) between individual groups as seen in (b) was carried out using one-way ANOVA with a Newman–Keul post hoc test from 3 sets of experimental results. Significance was assumed at ∗ P

    Article Snippet: Materials Etoposide was from Calbiochem.

    Techniques: SDS Page, Western Blot, Expressing

    (a) TCP80 has increased binding to the p53 mRNA following DNA damage. MCF-7 cells were transfected with pcDNA3.1/HisB/TCP80 that encodes for the Xpress-tagged TCP80 protein. Twenty-four hours following transfection, the cells were treated with or without 10 μ M etoposide for 2 hours. They were then lysed in polysome lysis buffer and incubated with protein G-plus agarose beads coated with the anti-Xpress antibody. The TCP80 and mRNA complexes (mRNP) were immunoprecipitated and mRNA was extracted from the immunoprecipitate as described in Experimental procedures. RT-PCR was then performed to reverse-transcribe and amplify the p53 IRES sequence (~145 bp). (b) TCP80 positively affects the p53 IRES activity in response to DNA damage. MCF-7 cells were cotransfected with pRF or pR5UTRF along with either pcDNA3.1 or pcDNA3.1/HisB/TCP80. Twenty-four hours following the transfection, the cells were treated with or without etoposide for 2 hours. The cells were then lysed and a dual-luciferase assay was performed to detect firefly (Fluc) and renilla (Rluc) luciferase activities as described in Experimental procedures. The results presented are average ± SEM from three individual experiments.

    Journal: BioMed Research International

    Article Title: Translational Control Protein 80 Stimulates IRES-Mediated Translation of p53 mRNA in Response to DNA Damage

    doi: 10.1155/2015/708158

    Figure Lengend Snippet: (a) TCP80 has increased binding to the p53 mRNA following DNA damage. MCF-7 cells were transfected with pcDNA3.1/HisB/TCP80 that encodes for the Xpress-tagged TCP80 protein. Twenty-four hours following transfection, the cells were treated with or without 10 μ M etoposide for 2 hours. They were then lysed in polysome lysis buffer and incubated with protein G-plus agarose beads coated with the anti-Xpress antibody. The TCP80 and mRNA complexes (mRNP) were immunoprecipitated and mRNA was extracted from the immunoprecipitate as described in Experimental procedures. RT-PCR was then performed to reverse-transcribe and amplify the p53 IRES sequence (~145 bp). (b) TCP80 positively affects the p53 IRES activity in response to DNA damage. MCF-7 cells were cotransfected with pRF or pR5UTRF along with either pcDNA3.1 or pcDNA3.1/HisB/TCP80. Twenty-four hours following the transfection, the cells were treated with or without etoposide for 2 hours. The cells were then lysed and a dual-luciferase assay was performed to detect firefly (Fluc) and renilla (Rluc) luciferase activities as described in Experimental procedures. The results presented are average ± SEM from three individual experiments.

    Article Snippet: Materials Etoposide was from Calbiochem.

    Techniques: Binding Assay, Transfection, Lysis, Incubation, Immunoprecipitation, Reverse Transcription Polymerase Chain Reaction, Sequencing, Activity Assay, Luciferase

    The results of the MTS-based assay of cytotoxicity of sulfoxide 2a in the concentration range from 18 to 2.25 mM. Control is presented by 1.5% solution of ethyl alcohol. Cytostatic compound of comparison is etoposide (1.25 mM, Calbiochem, USA).

    Journal: Frontiers in Pharmacology

    Article Title: Sulfur-Containing Monoterpenoids as Potential Antithrombotic Drugs: Research in the Molecular Mechanism of Coagulation Activity Using Pinanyl Sulfoxide as an Example

    doi: 10.3389/fphar.2018.00116

    Figure Lengend Snippet: The results of the MTS-based assay of cytotoxicity of sulfoxide 2a in the concentration range from 18 to 2.25 mM. Control is presented by 1.5% solution of ethyl alcohol. Cytostatic compound of comparison is etoposide (1.25 mM, Calbiochem, USA).

    Article Snippet: Briefly, human BJ fibroblasts (ATCC, USA) were seeded in 96-well flat-bottomed plates (Corning Inc., Corning NY) and allowed to attach and grow for 24 h in complete medium DMEM/199 supplemented with 10% of fetal veal serum (FBS, GIBCO, GrandIsland, NY), L-glutamine (0,3 mg/ml) and antibiotics penicillin-streptomycin (Paneko, Russia), The cells were cultured at 37°C of 5% of CO2 (LamSystems, Russia) with the indicated concentrations of pinanylsulfoxide 2a, 1.5% solution of ethyl alcohol or chemotherapeutic agent etoposide (Calbiochem, USA) (control).

    Techniques: MTS Assay, Concentration Assay

    Etoposide induced p53 activity is dampened in chronic hypoxia. D283-MED cells were incubated in 1% O 2 or 21% O 2 for 1 day or 5 days, prior to etoposide treatment where indicated. Three p53 target genes, MDM2, PUMA and p21 were assessed by qPCR. ( a ) Basal levels of p53 target genes in hypoxia without etoposide treatment were measured. The mRNA levels were normalised with the house-keeping gene cyclophilin A. ( b ) Levels of MDM2, p21 and PUMA mRNA with or without etoposide treatment. Data represented as normalised to housekeeping gene (cyclophilin A) and fold change with respect to the untreated control. Data are representative of three independent experiments, error bars are SD of a single experiment. ( c ) Total p53 and phosphorylated p53 serine 15 levels assessed by western blot. ( d ) Densitometry quantification of the band intensity was analysed by ImageJ from 3 independent experiments. Plot represents p53 serine 15 over the p53 total

    Journal: BMC Cancer

    Article Title: Decrease of Nibrin expression in chronic hypoxia is associated with hypoxia-induced chemoresistance in some brain tumour cells

    doi: 10.1186/s12885-019-5476-9

    Figure Lengend Snippet: Etoposide induced p53 activity is dampened in chronic hypoxia. D283-MED cells were incubated in 1% O 2 or 21% O 2 for 1 day or 5 days, prior to etoposide treatment where indicated. Three p53 target genes, MDM2, PUMA and p21 were assessed by qPCR. ( a ) Basal levels of p53 target genes in hypoxia without etoposide treatment were measured. The mRNA levels were normalised with the house-keeping gene cyclophilin A. ( b ) Levels of MDM2, p21 and PUMA mRNA with or without etoposide treatment. Data represented as normalised to housekeeping gene (cyclophilin A) and fold change with respect to the untreated control. Data are representative of three independent experiments, error bars are SD of a single experiment. ( c ) Total p53 and phosphorylated p53 serine 15 levels assessed by western blot. ( d ) Densitometry quantification of the band intensity was analysed by ImageJ from 3 independent experiments. Plot represents p53 serine 15 over the p53 total

    Article Snippet: Reagents Etoposide (E1383) was from Sigma.

    Techniques: Activity Assay, Incubation, Real-time Polymerase Chain Reaction, Western Blot

    Chronic hypoxia reduces ATM activation after etoposide treatment. D283-MED cells were pre-incubated in 21% O 2 , 1% O 2 or 0.1% O 2 prior to treatment with 20 μM etoposide for 4 h, or cells were left untreated as a control. Levels of ATM and ATM serine 1981 phosphorylation were determined using 3 independent western blots. The plots represent the densitometry of a representative western blot ( a ) measured using Image J

    Journal: BMC Cancer

    Article Title: Decrease of Nibrin expression in chronic hypoxia is associated with hypoxia-induced chemoresistance in some brain tumour cells

    doi: 10.1186/s12885-019-5476-9

    Figure Lengend Snippet: Chronic hypoxia reduces ATM activation after etoposide treatment. D283-MED cells were pre-incubated in 21% O 2 , 1% O 2 or 0.1% O 2 prior to treatment with 20 μM etoposide for 4 h, or cells were left untreated as a control. Levels of ATM and ATM serine 1981 phosphorylation were determined using 3 independent western blots. The plots represent the densitometry of a representative western blot ( a ) measured using Image J

    Article Snippet: Reagents Etoposide (E1383) was from Sigma.

    Techniques: Activation Assay, Incubation, Western Blot

    Etoposide efficacy is not affected by hypoxia. D283-MED cells were incubated in 1% O 2 for 5 days or in 21% O 2 prior to etoposide (20 μM) treatment for indicated time points. ( a ) ɣH2AX bound secondary Cy3 antibodies were detected by immunofluorescence, staining intensity was quantified for individual cells using AQM analysis. Data shown are the mean ± S.E.M of three independent experiments ( n > 350 cells). One-way ANOVA followed by Bonferroni test was performed (*indicates p

    Journal: BMC Cancer

    Article Title: Decrease of Nibrin expression in chronic hypoxia is associated with hypoxia-induced chemoresistance in some brain tumour cells

    doi: 10.1186/s12885-019-5476-9

    Figure Lengend Snippet: Etoposide efficacy is not affected by hypoxia. D283-MED cells were incubated in 1% O 2 for 5 days or in 21% O 2 prior to etoposide (20 μM) treatment for indicated time points. ( a ) ɣH2AX bound secondary Cy3 antibodies were detected by immunofluorescence, staining intensity was quantified for individual cells using AQM analysis. Data shown are the mean ± S.E.M of three independent experiments ( n > 350 cells). One-way ANOVA followed by Bonferroni test was performed (*indicates p

    Article Snippet: Reagents Etoposide (E1383) was from Sigma.

    Techniques: Incubation, Immunofluorescence, Staining

    Chronic hypoxia-induces treatment resistance in MB and glioblastoma cells. Cell viability was determined using an MTS assay, with absorbance normalised to untreated control. Two MB and one glioblastoma cell lines were pre-cultured in normoxia (21% O 2 ) or hypoxia (1% O 2 ) for the indicated time points prior to etoposide, cisplatin or X-ray irradiation treatment. Cells were maintained in 1% O 2 during etoposide [20–100 μM] and cisplatin [1–50 μg/ml] treatment. ( a ) D283-MED cells treated with etoposide ( b ) MEB-Med8A treated with etoposide ( c ) U87MG cells treated with etoposide. ( d ) D283-MED treated with cisplatin (1–5 μg/ml). ( e ) MEB-Med8A treated with cisplatin (1–5 μg/ml) ( f ) U87MG treated with cisplatin (30–50 μg/ml) ( g ) X-ray irradiation treatment was conducted in 21% O 2 with doses of 30 Gy (D283-MED), 50 Gy (MEB-Med8A) and 2 × 80 Gy dose (U87MG), with 48 h incubation post-treatment. The different doses is to account for different cell sensitivity to irradiation. Data are shown as the mean ± S.E.M of at least 3 independent experiments. A student t-test was performed where (*) indicates statistical significance with p

    Journal: BMC Cancer

    Article Title: Decrease of Nibrin expression in chronic hypoxia is associated with hypoxia-induced chemoresistance in some brain tumour cells

    doi: 10.1186/s12885-019-5476-9

    Figure Lengend Snippet: Chronic hypoxia-induces treatment resistance in MB and glioblastoma cells. Cell viability was determined using an MTS assay, with absorbance normalised to untreated control. Two MB and one glioblastoma cell lines were pre-cultured in normoxia (21% O 2 ) or hypoxia (1% O 2 ) for the indicated time points prior to etoposide, cisplatin or X-ray irradiation treatment. Cells were maintained in 1% O 2 during etoposide [20–100 μM] and cisplatin [1–50 μg/ml] treatment. ( a ) D283-MED cells treated with etoposide ( b ) MEB-Med8A treated with etoposide ( c ) U87MG cells treated with etoposide. ( d ) D283-MED treated with cisplatin (1–5 μg/ml). ( e ) MEB-Med8A treated with cisplatin (1–5 μg/ml) ( f ) U87MG treated with cisplatin (30–50 μg/ml) ( g ) X-ray irradiation treatment was conducted in 21% O 2 with doses of 30 Gy (D283-MED), 50 Gy (MEB-Med8A) and 2 × 80 Gy dose (U87MG), with 48 h incubation post-treatment. The different doses is to account for different cell sensitivity to irradiation. Data are shown as the mean ± S.E.M of at least 3 independent experiments. A student t-test was performed where (*) indicates statistical significance with p

    Article Snippet: Reagents Etoposide (E1383) was from Sigma.

    Techniques: MTS Assay, Cell Culture, Irradiation, Incubation

    Characterization of ultraviolet (UV) and etoposide (ETOP) induced DNA damage response (DDR). ( A ) DU145 cells were treated with UV at the indicated energy levels and cultured for 6 h. Western blot was performed for the indicated proteins. p-DNAPK: phosphorylation of DNAPK at serine 2056 (S2056); p-CHK1: phosphorylation of CHK1 at S345; p-CHK2: phosphorylation of CHK2 at threonine 68 (T68); ( B ) DU145 cells were treated with ETOP for 2 h at the indicated doses, followed by Western blot examination for the indicated proteins. All experiments were repeated once; typical images from a single repeat are shown.

    Journal: International Journal of Molecular Sciences

    Article Title: Microvesicles Contribute to the Bystander Effect of DNA Damage

    doi: 10.3390/ijms18040788

    Figure Lengend Snippet: Characterization of ultraviolet (UV) and etoposide (ETOP) induced DNA damage response (DDR). ( A ) DU145 cells were treated with UV at the indicated energy levels and cultured for 6 h. Western blot was performed for the indicated proteins. p-DNAPK: phosphorylation of DNAPK at serine 2056 (S2056); p-CHK1: phosphorylation of CHK1 at S345; p-CHK2: phosphorylation of CHK2 at threonine 68 (T68); ( B ) DU145 cells were treated with ETOP for 2 h at the indicated doses, followed by Western blot examination for the indicated proteins. All experiments were repeated once; typical images from a single repeat are shown.

    Article Snippet: Chemicals, Cell Lines, and Plasmids Hydroxyurea (HU) and etoposide (ETOP) were purchased from Sigma (Oakville, ON, Canada).

    Techniques: Cell Culture, Western Blot

    Etoposide and actinomycin D induced A3 expression and MT-COI editing in P2 cells. ( a ) Transcription profiling of A3A-A3H in etoposide or actinomycin D treated-P2 cells. Data in triplicate were normalized to the expression levels of RPL13A housekeeping reference genes and to untreated P2 cells to facilitate comparison (*p

    Journal: Scientific Reports

    Article Title: Genotoxic stress increases cytoplasmic mitochondrial DNA editing by human APOBEC3 mutator enzymes at a single cell level

    doi: 10.1038/s41598-019-39245-8

    Figure Lengend Snippet: Etoposide and actinomycin D induced A3 expression and MT-COI editing in P2 cells. ( a ) Transcription profiling of A3A-A3H in etoposide or actinomycin D treated-P2 cells. Data in triplicate were normalized to the expression levels of RPL13A housekeeping reference genes and to untreated P2 cells to facilitate comparison (*p

    Article Snippet: Reagents and Plasmids Etoposide was from Sigma and actinomycin D from Millipore.

    Techniques: Expressing

    Frequencies of A3-edited cymtDNA in single cells. Frequencies of single cells harboring A3-edited cymtDNA using a fixed PCR denaturation temperature of 85 °C. Analysis was performed in single CD4 + T lymphocytes from 2 donors D1 and D2, ~17% and 12% of cells scored positive for APOBEC3 edited cymtDNA. Among the 512 P2 ung − / − cells analyzed at 85 °C, 3 cells (0.6%) are positive. P2 ung − / − cells were treated with 100 μM actinomycin D (act D) or etoposide (etop) for 16 hours. Treatment increased the proportion of cells showing evidence of cymtDNA editing to ~24.6% and 22.2% with actinomycin D or etoposide respectively. f: frequency, #: number.

    Journal: Scientific Reports

    Article Title: Genotoxic stress increases cytoplasmic mitochondrial DNA editing by human APOBEC3 mutator enzymes at a single cell level

    doi: 10.1038/s41598-019-39245-8

    Figure Lengend Snippet: Frequencies of A3-edited cymtDNA in single cells. Frequencies of single cells harboring A3-edited cymtDNA using a fixed PCR denaturation temperature of 85 °C. Analysis was performed in single CD4 + T lymphocytes from 2 donors D1 and D2, ~17% and 12% of cells scored positive for APOBEC3 edited cymtDNA. Among the 512 P2 ung − / − cells analyzed at 85 °C, 3 cells (0.6%) are positive. P2 ung − / − cells were treated with 100 μM actinomycin D (act D) or etoposide (etop) for 16 hours. Treatment increased the proportion of cells showing evidence of cymtDNA editing to ~24.6% and 22.2% with actinomycin D or etoposide respectively. f: frequency, #: number.

    Article Snippet: Reagents and Plasmids Etoposide was from Sigma and actinomycin D from Millipore.

    Techniques: Polymerase Chain Reaction, Activated Clotting Time Assay

    Bid-overexpression on the activation of Akt in response to etoposide. Vector control ( A ) and Bid/PLC/PRF/5 cells ( B ) were treated with etoposide for different periods, respectively. Then, p-Akt was detected by Western blot analysis. All blots were subsequently stripped and reprobed with antibodies against Akt. The density of pSer 473 -Akt protein bands was determined ( C ). Abbreviation: Bid, BH3-interacting domain death agonist.

    Journal: OncoTargets and therapy

    Article Title: Bid-overexpression regulates proliferation and phosphorylation of Akt and MAPKs in response to etoposide-induced DNA damage in hepatocellular carcinoma cells

    doi: 10.2147/OTT.S36087

    Figure Lengend Snippet: Bid-overexpression on the activation of Akt in response to etoposide. Vector control ( A ) and Bid/PLC/PRF/5 cells ( B ) were treated with etoposide for different periods, respectively. Then, p-Akt was detected by Western blot analysis. All blots were subsequently stripped and reprobed with antibodies against Akt. The density of pSer 473 -Akt protein bands was determined ( C ). Abbreviation: Bid, BH3-interacting domain death agonist.

    Article Snippet: Reagents and antibodies Etoposide was purchased from Calbiochem (USA).

    Techniques: Over Expression, Activation Assay, Plasmid Preparation, Planar Chromatography, Western Blot

    Effects of Bid-overexpression on the phosphorylation of ERK1/2 in response to etoposide. Vector control ( A ) and Bid/PLC/PRF/5 cells ( B ) were treated with etoposide for different periods, respectively. Then p-ERK1/2 was detected by Western blot analysis. All blots were subsequently stripped and reprobed with antibodies against ERK1/2. The density of p-ERK1/2 protein bands was determined ( C ). Abbreviation: Bid, BH3-interacting domain death agonist.

    Journal: OncoTargets and therapy

    Article Title: Bid-overexpression regulates proliferation and phosphorylation of Akt and MAPKs in response to etoposide-induced DNA damage in hepatocellular carcinoma cells

    doi: 10.2147/OTT.S36087

    Figure Lengend Snippet: Effects of Bid-overexpression on the phosphorylation of ERK1/2 in response to etoposide. Vector control ( A ) and Bid/PLC/PRF/5 cells ( B ) were treated with etoposide for different periods, respectively. Then p-ERK1/2 was detected by Western blot analysis. All blots were subsequently stripped and reprobed with antibodies against ERK1/2. The density of p-ERK1/2 protein bands was determined ( C ). Abbreviation: Bid, BH3-interacting domain death agonist.

    Article Snippet: Reagents and antibodies Etoposide was purchased from Calbiochem (USA).

    Techniques: Over Expression, Plasmid Preparation, Planar Chromatography, Western Blot

    Effects of Bid-overexpression on the phosphorylation of c-Jun in response to etoposide. Vector control ( A ) and Bid/PLC/PRF/5 cells ( B ) were treated with etoposide for different periods, respectively. Then, p-c-Jun was detected by Western blot analysis. All blots were subsequently stripped and reprobed with antibodies against c-Jun. The density of p-c-Jun protein bands was determined ( C ). Abbreviation: Bid, BH3-interacting domain death agonist.

    Journal: OncoTargets and therapy

    Article Title: Bid-overexpression regulates proliferation and phosphorylation of Akt and MAPKs in response to etoposide-induced DNA damage in hepatocellular carcinoma cells

    doi: 10.2147/OTT.S36087

    Figure Lengend Snippet: Effects of Bid-overexpression on the phosphorylation of c-Jun in response to etoposide. Vector control ( A ) and Bid/PLC/PRF/5 cells ( B ) were treated with etoposide for different periods, respectively. Then, p-c-Jun was detected by Western blot analysis. All blots were subsequently stripped and reprobed with antibodies against c-Jun. The density of p-c-Jun protein bands was determined ( C ). Abbreviation: Bid, BH3-interacting domain death agonist.

    Article Snippet: Reagents and antibodies Etoposide was purchased from Calbiochem (USA).

    Techniques: Over Expression, Plasmid Preparation, Planar Chromatography, Western Blot

    Effect of Bid on the PLC/PRF/5 cell viability in the response of etoposide-induced DNA damage. Overexpression of Bid does not alter the rate of cell proliferation under low concentration of etoposide, but Bid-overexpression cells sensitize apoptosis induced by high concentration of etoposide. ( A and B ) Cell viability was determined by the MTT assay method. ( C ) Cell proliferation rate was determined by the BrdU-labeling method assay. Notes: Data are reported as means ± standard deviations of three independent experiments; *indicates P

    Journal: OncoTargets and therapy

    Article Title: Bid-overexpression regulates proliferation and phosphorylation of Akt and MAPKs in response to etoposide-induced DNA damage in hepatocellular carcinoma cells

    doi: 10.2147/OTT.S36087

    Figure Lengend Snippet: Effect of Bid on the PLC/PRF/5 cell viability in the response of etoposide-induced DNA damage. Overexpression of Bid does not alter the rate of cell proliferation under low concentration of etoposide, but Bid-overexpression cells sensitize apoptosis induced by high concentration of etoposide. ( A and B ) Cell viability was determined by the MTT assay method. ( C ) Cell proliferation rate was determined by the BrdU-labeling method assay. Notes: Data are reported as means ± standard deviations of three independent experiments; *indicates P

    Article Snippet: Reagents and antibodies Etoposide was purchased from Calbiochem (USA).

    Techniques: Planar Chromatography, Over Expression, Concentration Assay, MTT Assay, Labeling

    Effects of Bid-overexpression on the phosphorylation of p38 in response to etoposide. Vector control ( A ) and Bid/PLC/PRF/5 cells ( B ) were treated with etoposide for different periods of time, respectively. Then p-p38 was detected by Western blot analysis. All blots were subsequently stripped and reprobed with antibodies against p38. The density of p-p38 protein bands was determined ( C ). Abbreviation: Bid, BH3-interacting domain death agonist.

    Journal: OncoTargets and therapy

    Article Title: Bid-overexpression regulates proliferation and phosphorylation of Akt and MAPKs in response to etoposide-induced DNA damage in hepatocellular carcinoma cells

    doi: 10.2147/OTT.S36087

    Figure Lengend Snippet: Effects of Bid-overexpression on the phosphorylation of p38 in response to etoposide. Vector control ( A ) and Bid/PLC/PRF/5 cells ( B ) were treated with etoposide for different periods of time, respectively. Then p-p38 was detected by Western blot analysis. All blots were subsequently stripped and reprobed with antibodies against p38. The density of p-p38 protein bands was determined ( C ). Abbreviation: Bid, BH3-interacting domain death agonist.

    Article Snippet: Reagents and antibodies Etoposide was purchased from Calbiochem (USA).

    Techniques: Over Expression, Plasmid Preparation, Planar Chromatography, Western Blot

    Different apoptotic stimuli results in the cleavage of K18 and EndoB. ( A ) Western blot analysis of HR-9 (lanes 1–7 ) and SNG-M (lanes 8–14 ) cells subjected to different apoptotic stimuli. Adherent ( A ) and floating ( F ) cells were separately collected after etoposide ( ETP ) and daunomycin ( DNM ) treatment and UV exposure, and whole-cell lysates were analyzed by Western blot with polyclonal anti-K18. Lanes 1 and 9 represent control untreated cells. ( B ) Low molecular weight genomic DNA was analyzed from cells represented in A except for lane 1 , which contained size markers. ( C ) Comparison of K18 found in apoptotic cells and purified K18 cleaved in vitro by caspase-6. Cell lysates or purified protein preparation was analyzed by Western blotting with antibody 3055. Untreated SNG-M cells (lane 1 ), etoposide ( ETP ) treated floating SNG-M cells (lane 7 ), and mouse HR-9 cells (lane 2 ) or apoptotic HR-9 cells (lane 6 ) were compared with purified K18 (lane 4 ), purified K18 mixed with HR-9 cell lysate (lane 3 ), or K18 cleaved with caspase-6 ( C6 ) in vitro and mixed with HR-9 cell lysate (lane 5 ). Note the comigration of caspase-6–cleaved K18 (lane 5 ) and K18 B found in apoptotic SNG-M cells (lane 7 ).

    Journal: The Journal of Cell Biology

    Article Title: Caspase Cleavage of Keratin 18 and Reorganization of Intermediate Filaments during Epithelial Cell Apoptosis

    doi:

    Figure Lengend Snippet: Different apoptotic stimuli results in the cleavage of K18 and EndoB. ( A ) Western blot analysis of HR-9 (lanes 1–7 ) and SNG-M (lanes 8–14 ) cells subjected to different apoptotic stimuli. Adherent ( A ) and floating ( F ) cells were separately collected after etoposide ( ETP ) and daunomycin ( DNM ) treatment and UV exposure, and whole-cell lysates were analyzed by Western blot with polyclonal anti-K18. Lanes 1 and 9 represent control untreated cells. ( B ) Low molecular weight genomic DNA was analyzed from cells represented in A except for lane 1 , which contained size markers. ( C ) Comparison of K18 found in apoptotic cells and purified K18 cleaved in vitro by caspase-6. Cell lysates or purified protein preparation was analyzed by Western blotting with antibody 3055. Untreated SNG-M cells (lane 1 ), etoposide ( ETP ) treated floating SNG-M cells (lane 7 ), and mouse HR-9 cells (lane 2 ) or apoptotic HR-9 cells (lane 6 ) were compared with purified K18 (lane 4 ), purified K18 mixed with HR-9 cell lysate (lane 3 ), or K18 cleaved with caspase-6 ( C6 ) in vitro and mixed with HR-9 cell lysate (lane 5 ). Note the comigration of caspase-6–cleaved K18 (lane 5 ) and K18 B found in apoptotic SNG-M cells (lane 7 ).

    Article Snippet: Apoptosis was also induced by treatment of semiconfluent cultures with the drugs etoposide ( Sigma Chemical Co. ) at 250 μg/ml for SNG-M and 125 μg/ml for HR-9 and daunomycin ( Sigma Chemical Co. ) at 6 μg/ml for SNG-M and 2 μg/ml for HR-9.

    Techniques: Western Blot, Molecular Weight, Purification, In Vitro

    K18 granular structures are preferentially phosphorylated. Etoposide-treated SNG-M cells were fixed and stained by indirect immunofluorescence for phospho-ser53 K18 with antibody 3055 ( A , C , and E ) and with propidium iodide for DNA ( B , D , and F ). Note the granular keratin staining of some cells with normal nuclear morphology in A , the staining of cell–cell junctions in A and C , and advanced keratin disorganization with nuclear fragmentation in C and E.

    Journal: The Journal of Cell Biology

    Article Title: Caspase Cleavage of Keratin 18 and Reorganization of Intermediate Filaments during Epithelial Cell Apoptosis

    doi:

    Figure Lengend Snippet: K18 granular structures are preferentially phosphorylated. Etoposide-treated SNG-M cells were fixed and stained by indirect immunofluorescence for phospho-ser53 K18 with antibody 3055 ( A , C , and E ) and with propidium iodide for DNA ( B , D , and F ). Note the granular keratin staining of some cells with normal nuclear morphology in A , the staining of cell–cell junctions in A and C , and advanced keratin disorganization with nuclear fragmentation in C and E.

    Article Snippet: Apoptosis was also induced by treatment of semiconfluent cultures with the drugs etoposide ( Sigma Chemical Co. ) at 250 μg/ml for SNG-M and 125 μg/ml for HR-9 and daunomycin ( Sigma Chemical Co. ) at 6 μg/ml for SNG-M and 2 μg/ml for HR-9.

    Techniques: Staining, Immunofluorescence

    Treatment of SNG-M cells with etoposide results in specific cleavage of K18. ( A ) SNG-M cells were treated with etoposide (250 μg/ml) for 0, 12, 24, 36, 48, 60, 72, and 84 h. Adherent and floating cells were combined, and whole-cell lysates were analyzed by Western blot with polyclonal antibody anti-K18 1589. This membrane was stripped and probed consecutively with monoclonal anti-K8 M20 ( C ), anti–phospho ser53-K18 antibodies ( B ), and monoclonal anti-K18 CK5 ( D ). ( E ) HR-9 cells were treated with daunomycin for 0, 12, 24, 36, 48, and 60 h. Adherent and floating cells were combined, and whole-cell lysates were analyzed by Western blot with anti-K18 polyclonal antibody, 1589.

    Journal: The Journal of Cell Biology

    Article Title: Caspase Cleavage of Keratin 18 and Reorganization of Intermediate Filaments during Epithelial Cell Apoptosis

    doi:

    Figure Lengend Snippet: Treatment of SNG-M cells with etoposide results in specific cleavage of K18. ( A ) SNG-M cells were treated with etoposide (250 μg/ml) for 0, 12, 24, 36, 48, 60, 72, and 84 h. Adherent and floating cells were combined, and whole-cell lysates were analyzed by Western blot with polyclonal antibody anti-K18 1589. This membrane was stripped and probed consecutively with monoclonal anti-K8 M20 ( C ), anti–phospho ser53-K18 antibodies ( B ), and monoclonal anti-K18 CK5 ( D ). ( E ) HR-9 cells were treated with daunomycin for 0, 12, 24, 36, 48, and 60 h. Adherent and floating cells were combined, and whole-cell lysates were analyzed by Western blot with anti-K18 polyclonal antibody, 1589.

    Article Snippet: Apoptosis was also induced by treatment of semiconfluent cultures with the drugs etoposide ( Sigma Chemical Co. ) at 250 μg/ml for SNG-M and 125 μg/ml for HR-9 and daunomycin ( Sigma Chemical Co. ) at 6 μg/ml for SNG-M and 2 μg/ml for HR-9.

    Techniques: Western Blot

    ( A ) Kinetics of induction of apoptosis in etoposide-treated SNG-M cells. In parallel experiments to the ones described in Fig. 3 A , SNG-M cells were treated with etoposide (250 μg/ml) for 0, 12, 24, 36, 48, 60, 72, and 84 h. Adherent and floating cells were combined, fixed in methanol, and stained with DAPI. Nuclei fragmentation was used as the criterion of apoptosis. ( B ) Comparison of the kinetics of appearance of the K18 23-kD fragment (K18 B) and its phosphorylated form (P-K18 B) by densitometric analysis of the signal detected in Western blotting shown in Fig. 3 , A and B. The y-axis represents the percentage of the maximum signal. Maximum signal for P-K18 B was obtained after 36 h and for K18 B after 60 h. ( C ) Comparison of the kinetics of disappearance of K18 and P-K18 by densitometric analysis of the signal detected in Fig. 3 , A and B.

    Journal: The Journal of Cell Biology

    Article Title: Caspase Cleavage of Keratin 18 and Reorganization of Intermediate Filaments during Epithelial Cell Apoptosis

    doi:

    Figure Lengend Snippet: ( A ) Kinetics of induction of apoptosis in etoposide-treated SNG-M cells. In parallel experiments to the ones described in Fig. 3 A , SNG-M cells were treated with etoposide (250 μg/ml) for 0, 12, 24, 36, 48, 60, 72, and 84 h. Adherent and floating cells were combined, fixed in methanol, and stained with DAPI. Nuclei fragmentation was used as the criterion of apoptosis. ( B ) Comparison of the kinetics of appearance of the K18 23-kD fragment (K18 B) and its phosphorylated form (P-K18 B) by densitometric analysis of the signal detected in Western blotting shown in Fig. 3 , A and B. The y-axis represents the percentage of the maximum signal. Maximum signal for P-K18 B was obtained after 36 h and for K18 B after 60 h. ( C ) Comparison of the kinetics of disappearance of K18 and P-K18 by densitometric analysis of the signal detected in Fig. 3 , A and B.

    Article Snippet: Apoptosis was also induced by treatment of semiconfluent cultures with the drugs etoposide ( Sigma Chemical Co. ) at 250 μg/ml for SNG-M and 125 μg/ml for HR-9 and daunomycin ( Sigma Chemical Co. ) at 6 μg/ml for SNG-M and 2 μg/ml for HR-9.

    Techniques: Staining, Western Blot

    Induction of apoptosis in SNG-M cells results in reorganization of K18 IF. SNG-M cells were treated with etoposide (250 μg/ml) for 24 h and stained by indirect immunofluorescence for K18 with monoclonal antibody CK5 ( A , C , and E ) and with propidium iodide for DNA ( B , D , and F ). Images were analyzed with a confocal microscope. Note the granular K18 staining associated with altered nuclear morphology.

    Journal: The Journal of Cell Biology

    Article Title: Caspase Cleavage of Keratin 18 and Reorganization of Intermediate Filaments during Epithelial Cell Apoptosis

    doi:

    Figure Lengend Snippet: Induction of apoptosis in SNG-M cells results in reorganization of K18 IF. SNG-M cells were treated with etoposide (250 μg/ml) for 24 h and stained by indirect immunofluorescence for K18 with monoclonal antibody CK5 ( A , C , and E ) and with propidium iodide for DNA ( B , D , and F ). Images were analyzed with a confocal microscope. Note the granular K18 staining associated with altered nuclear morphology.

    Article Snippet: Apoptosis was also induced by treatment of semiconfluent cultures with the drugs etoposide ( Sigma Chemical Co. ) at 250 μg/ml for SNG-M and 125 μg/ml for HR-9 and daunomycin ( Sigma Chemical Co. ) at 6 μg/ml for SNG-M and 2 μg/ml for HR-9.

    Techniques: Staining, Immunofluorescence, Microscopy

    Association between NUP153 dependency and cell cycle independence. ( A ) Propidium iodide staining of HOS cells untreated (grey) or treated for 24 h with 5 µM Etoposide phosphate (red line). ( B ) Infectivity of CA mutant viruses in HOS cells arrested with 5 µM Etoposide phosphate, normalized to infectivity in control HOS cells. Error bars denote standard error of 4 experiments. ( C ) Scatter plot comparison of CA mutant sensitivities in cell cycle arrested HOS cells in the absence or presence of 5 µM cyclosporine (CsA). Mutant viruses most sensitive to cell cycle arrest are indicated. ( D to G ) Scatter plots comparing sensitivities of mutant viruses to cell cycle arrest versus NUP153 knockdown (D), or restriction by Trim-NUP153 C (E), CPSF6 358 (F), or Trim-CPSF6 358 (G). Spearman rank correlation coefficients and measures of significance are indicated. Data points for CA mutant viruses P38A, T54A, A92E, and G94D clustered with the WT virus within these panels, so their labels were omitted to aid legibility.

    Journal: PLoS Pathogens

    Article Title: Nucleoporin NUP153 Phenylalanine-Glycine Motifs Engage a Common Binding Pocket within the HIV-1 Capsid Protein to Mediate Lentiviral Infectivity

    doi: 10.1371/journal.ppat.1003693

    Figure Lengend Snippet: Association between NUP153 dependency and cell cycle independence. ( A ) Propidium iodide staining of HOS cells untreated (grey) or treated for 24 h with 5 µM Etoposide phosphate (red line). ( B ) Infectivity of CA mutant viruses in HOS cells arrested with 5 µM Etoposide phosphate, normalized to infectivity in control HOS cells. Error bars denote standard error of 4 experiments. ( C ) Scatter plot comparison of CA mutant sensitivities in cell cycle arrested HOS cells in the absence or presence of 5 µM cyclosporine (CsA). Mutant viruses most sensitive to cell cycle arrest are indicated. ( D to G ) Scatter plots comparing sensitivities of mutant viruses to cell cycle arrest versus NUP153 knockdown (D), or restriction by Trim-NUP153 C (E), CPSF6 358 (F), or Trim-CPSF6 358 (G). Spearman rank correlation coefficients and measures of significance are indicated. Data points for CA mutant viruses P38A, T54A, A92E, and G94D clustered with the WT virus within these panels, so their labels were omitted to aid legibility.

    Article Snippet: Cell cycle arrest experiments were performed by plating 2,500 control or 5,000 experimental cells treated with 5 µM Etoposide-phosphate (Calbiochem) the day before infection.

    Techniques: Staining, Infection, Mutagenesis

    miR-3196 inhibits VP-16 induced apoptosis by downregulating PUMA A. A549 cells were transfected with miR-3196 mimics (miR-3196) and/or combined with pcDNA3-PUMA (PUMA) for 24 h, followed by treatment with VP16 (100 μM) for 48 h. The apoptotic cells were detected by FCM (left panel). Error bars denote the mean ± SD (right panel). B. H1650 cells were transfected and treated as in (A). The apoptotic cells were detected by FCM (left panel). Error bars denote the mean ± SD (right panel). * P

    Journal: Oncotarget

    Article Title: MicroRNA-3196 is inhibited by H2AX phosphorylation and attenuates lung cancer cell apoptosis by downregulating PUMA

    doi: 10.18632/oncotarget.12794

    Figure Lengend Snippet: miR-3196 inhibits VP-16 induced apoptosis by downregulating PUMA A. A549 cells were transfected with miR-3196 mimics (miR-3196) and/or combined with pcDNA3-PUMA (PUMA) for 24 h, followed by treatment with VP16 (100 μM) for 48 h. The apoptotic cells were detected by FCM (left panel). Error bars denote the mean ± SD (right panel). B. H1650 cells were transfected and treated as in (A). The apoptotic cells were detected by FCM (left panel). Error bars denote the mean ± SD (right panel). * P

    Article Snippet: Cells were grown to 80% confluence and then serum-deprived for 12 h prior to etoposide (VP16) (Sigma–Aldrich, St Louis, MO, USA) stimulation.

    Techniques: Transfection

    GDNF and VP-16 expression patterns . The GDNF expression pattern of the final ON group ( a , b ) was similar to that previously observed where the GDNF transgene (red) is observed distally from that of the transduced cells and VP-16 (green). ( b ) Increased magnification of area outlined in a . VP-16 expression in the OFF group ( c , d ) was similar to that seen in the ON group; however, no GDNF protein expression was observed. GDNF protein expression in the CBA-GDNF group ( e , f ) extended in a diffuse pattern into the VTA and the DMN indicating that GDNF protein was distributed away from the transduced cells in contrast to the pattern of expression observed from an intracellular transgene such as the SN-GFP group ( g , h ). As expected, no immunoreactivity was observed for VP-16 in either the CBA-GDNF group ( e , f , green) or the SN-GFP group ( g , h , red). ( f and h ) Higher magnifications of areas outlined in e and g , respectively. Bar: a , c , e , g = 1.0 mm, bar: b , d , f , h = 0.1 mm. CBA, chicken β-actin; DMN, deep mesencephalic nucleus; GDNF, glial cell line-derived neurotrophic factor; SNc, substantia nigra pars compacta; SNr, substantia nigra pars reticulate; VTA, ventral tegmental area.

    Journal: Molecular Therapy

    Article Title: Tight Long-term Dynamic Doxycycline Responsive Nigrostriatal GDNF Using a Single rAAV Vector

    doi: 10.1038/mt.2009.196

    Figure Lengend Snippet: GDNF and VP-16 expression patterns . The GDNF expression pattern of the final ON group ( a , b ) was similar to that previously observed where the GDNF transgene (red) is observed distally from that of the transduced cells and VP-16 (green). ( b ) Increased magnification of area outlined in a . VP-16 expression in the OFF group ( c , d ) was similar to that seen in the ON group; however, no GDNF protein expression was observed. GDNF protein expression in the CBA-GDNF group ( e , f ) extended in a diffuse pattern into the VTA and the DMN indicating that GDNF protein was distributed away from the transduced cells in contrast to the pattern of expression observed from an intracellular transgene such as the SN-GFP group ( g , h ). As expected, no immunoreactivity was observed for VP-16 in either the CBA-GDNF group ( e , f , green) or the SN-GFP group ( g , h , red). ( f and h ) Higher magnifications of areas outlined in e and g , respectively. Bar: a , c , e , g = 1.0 mm, bar: b , d , f , h = 0.1 mm. CBA, chicken β-actin; DMN, deep mesencephalic nucleus; GDNF, glial cell line-derived neurotrophic factor; SNc, substantia nigra pars compacta; SNr, substantia nigra pars reticulate; VTA, ventral tegmental area.

    Article Snippet: Primary antibodies used were mouse anti-TH (1:2,000), rabbit anti-GFP (1:2,000), mouse anti-calbindin (1:200), rabbit anti-VP-16 (1:50) (Chemicon, Temecula, CA), goat anti-GDNF (1:1,000) (R & D Systems, Minneapolis, MN).

    Techniques: Expressing, Crocin Bleaching Assay, Derivative Assay

    Experimental design and vector schematics . rAAV genomic maps of vectors used in all three experiments. ( a ) CBA-GDNF, ( b ) CBA-GFP, and ( c ) regGDNF. ( d ) Experimental design and timeline of the three experiments performed in this study. The alternating white gray areas indicate the crossover of feeding DOX or normal food in the ON–OFF experiment. At the end of the ON–OFF study, SN-regGDNF1 was in the OFF state (expressing GDNF) and SN-regGDNF2 was in the ON state. The different shades of gray in the dose–response experiment denote the fact that the DOX diet doses were changed during that experiment as indicated. No DOX was administered during the striatum experiment. ( e ) Photograph of the ventral surface of the rat brain displaying how the brain was divided for the assays and histology as described. CMVe, cytomegalovirus enhancer element; DOX, doxycycline; eGFP, enhanced green fluorescent protein; hGDNF, human glial cell line-derived neurotrophic factor; iTR, inverted terminal repeat; pCBA, chicken β-actin promoter; pTET-dCMV minimal , tetracycline-responsive delta-CMV minimal promoter; rAAV, recombinant adeno-associated virus; SV40 pA, SV40 poly-A sequence; tTA2, transactivator containing VP-16 sequence.

    Journal: Molecular Therapy

    Article Title: Tight Long-term Dynamic Doxycycline Responsive Nigrostriatal GDNF Using a Single rAAV Vector

    doi: 10.1038/mt.2009.196

    Figure Lengend Snippet: Experimental design and vector schematics . rAAV genomic maps of vectors used in all three experiments. ( a ) CBA-GDNF, ( b ) CBA-GFP, and ( c ) regGDNF. ( d ) Experimental design and timeline of the three experiments performed in this study. The alternating white gray areas indicate the crossover of feeding DOX or normal food in the ON–OFF experiment. At the end of the ON–OFF study, SN-regGDNF1 was in the OFF state (expressing GDNF) and SN-regGDNF2 was in the ON state. The different shades of gray in the dose–response experiment denote the fact that the DOX diet doses were changed during that experiment as indicated. No DOX was administered during the striatum experiment. ( e ) Photograph of the ventral surface of the rat brain displaying how the brain was divided for the assays and histology as described. CMVe, cytomegalovirus enhancer element; DOX, doxycycline; eGFP, enhanced green fluorescent protein; hGDNF, human glial cell line-derived neurotrophic factor; iTR, inverted terminal repeat; pCBA, chicken β-actin promoter; pTET-dCMV minimal , tetracycline-responsive delta-CMV minimal promoter; rAAV, recombinant adeno-associated virus; SV40 pA, SV40 poly-A sequence; tTA2, transactivator containing VP-16 sequence.

    Article Snippet: Primary antibodies used were mouse anti-TH (1:2,000), rabbit anti-GFP (1:2,000), mouse anti-calbindin (1:200), rabbit anti-VP-16 (1:50) (Chemicon, Temecula, CA), goat anti-GDNF (1:1,000) (R & D Systems, Minneapolis, MN).

    Techniques: Plasmid Preparation, Crocin Bleaching Assay, Expressing, Derivative Assay, Recombinant, Sequencing

    Effects of engraftment location, estrogen, and immune suppressant on tumorigenicity of breast cancer cells NOD/SCID mice were treated ± 17β-estradiol (E 2 ) and etoposide as indicated. Six days later, mice were surgically incised and injected with 500 cells from a breast cancer pleural effusion in 25% Matrigel. A: final tumor volume. Data represent mean ± SE. B: tumor incidence. C: latency of tumor formation following injection (* P

    Journal: Journal of cellular physiology

    Article Title: Local regulation of human breast xenograft models

    doi: 10.1002/jcp.22190

    Figure Lengend Snippet: Effects of engraftment location, estrogen, and immune suppressant on tumorigenicity of breast cancer cells NOD/SCID mice were treated ± 17β-estradiol (E 2 ) and etoposide as indicated. Six days later, mice were surgically incised and injected with 500 cells from a breast cancer pleural effusion in 25% Matrigel. A: final tumor volume. Data represent mean ± SE. B: tumor incidence. C: latency of tumor formation following injection (* P

    Article Snippet: Six days prior to cell injection, mice were treated ± subcutaneous 17β-estradiol pellet (0.72 mg, 90-day release; Innovative Research of America, Sarasota, FL) and ± the bone marrow suppressant VP-16 (etoposide, a commonly used immune suppressant, 0.6 mg; Calbiochem, Gibbstown, NJ) administered i.p. ( ; ; ).

    Techniques: Mouse Assay, Injection

    The combined use of anti-CD33 mAb and etoposide displays a synergistic effect in binding of Annexin V. The surface binding of Annexin V to AML cells was measured after 20 h of culture with GM-CSF in the presence of anti-CD33 mAb (μg/ml), ETP (68 μM/ml), or ETP (34 μM/ml) used either alone or in combination. The experiment shown is representative of four independent experiments. Samples were stained with Annexin V FITC-conjugated mAb. Analysis was performed on PI-negative, viable cells.

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

    Article Title: Surface expression and function of p75/AIRM-1 or CD33 in acute myeloid leukemias: Engagement of CD33 induces apoptosis of leukemic cells

    doi: 10.1073/pnas.091097198

    Figure Lengend Snippet: The combined use of anti-CD33 mAb and etoposide displays a synergistic effect in binding of Annexin V. The surface binding of Annexin V to AML cells was measured after 20 h of culture with GM-CSF in the presence of anti-CD33 mAb (μg/ml), ETP (68 μM/ml), or ETP (34 μM/ml) used either alone or in combination. The experiment shown is representative of four independent experiments. Samples were stained with Annexin V FITC-conjugated mAb. Analysis was performed on PI-negative, viable cells.

    Article Snippet: Etoposide-VP16 (ETP) was purchased from Sigma.

    Techniques: Binding Assay, Staining

    MMS22L foci form during S phase of the cell cycle and colocalize with HR proteins Immunofluorescence images of MMS22L foci formation at etoposide (ETP)‐induced DSBs during S, M, and G1 phases of the cell cycle in HeLa cells. Scale bar: 5 μm. Quantification of a representative experiment from (A) shows number of MMS22L foci per nucleus during indicated cell cycle phases ( n = 2; n nuclei ≥ 60). Immunofluorescence images of CPT‐induced MMS22L, RPA2, BRCA2, and RAD51 foci in U2OS cells. Scale bar: 5 μm. Quantification of a representative experiment ( n = 3) from (C) shows percentage of γH2AX, RPA2, BRCA2, and RAD51 foci colocalizing with MMS22L foci ( n nuclei = 30; nuclei with ≥ 25 MMS22L foci were analyzed). Data information: Boxplots in (B and D) represent distributions per nucleus; boxes indicate the 25–75 percentile and whiskers the 10–90 percentile. Horizontal lines mark the medians. Statistical analysis: Mann–Whitney U ‐test; *** P ≤ 0.0001; ns, not significant.

    Journal: The EMBO Journal

    Article Title: The MMS22L–TONSL heterodimer directly promotes RAD51‐dependent recombination upon replication stress

    doi: 10.15252/embj.201593132

    Figure Lengend Snippet: MMS22L foci form during S phase of the cell cycle and colocalize with HR proteins Immunofluorescence images of MMS22L foci formation at etoposide (ETP)‐induced DSBs during S, M, and G1 phases of the cell cycle in HeLa cells. Scale bar: 5 μm. Quantification of a representative experiment from (A) shows number of MMS22L foci per nucleus during indicated cell cycle phases ( n = 2; n nuclei ≥ 60). Immunofluorescence images of CPT‐induced MMS22L, RPA2, BRCA2, and RAD51 foci in U2OS cells. Scale bar: 5 μm. Quantification of a representative experiment ( n = 3) from (C) shows percentage of γH2AX, RPA2, BRCA2, and RAD51 foci colocalizing with MMS22L foci ( n nuclei = 30; nuclei with ≥ 25 MMS22L foci were analyzed). Data information: Boxplots in (B and D) represent distributions per nucleus; boxes indicate the 25–75 percentile and whiskers the 10–90 percentile. Horizontal lines mark the medians. Statistical analysis: Mann–Whitney U ‐test; *** P ≤ 0.0001; ns, not significant.

    Article Snippet: Cells were treated with the following drugs diluted in growth medium: Topo II inhibitor etoposide (5 μM; Sigma‐Aldrich), Topo I inhibitor camptothecin (Sigma‐Aldrich), hydroxyurea (Sigma‐Aldrich), polymerase‐alpha inhibitor aphidicolin (2 μM; Sigma‐Aldrich), and ATR inhibitor VE‐821 (3 μM; Selleck Chemicals).

    Techniques: Immunofluorescence, Cycling Probe Technology, MANN-WHITNEY

    Flow cytometry analysis of the rates of apoptosis in Huh7 cells exposed to DIC or to IND . For apoptosis analysis, permeabilized Huh7 cells were labelled with propidium iodide (PI) and analysed by flow cytometry by a FACSCalibur flow cytometer (Becton Dickinson, North Ryde, NSW, Australia). DNA damaging agent etopoxide was used as positive control for apoptosis. Results are the mean of three experiments repeated three times.

    Journal: The Open Biochemistry Journal

    Article Title: In the Huh7 Hepatoma Cells Diclofenac and Indomethacin Activate Differently the Unfolded Protein Response and Induce ER Stress Apoptosis

    doi: 10.2174/1874091X01105010045

    Figure Lengend Snippet: Flow cytometry analysis of the rates of apoptosis in Huh7 cells exposed to DIC or to IND . For apoptosis analysis, permeabilized Huh7 cells were labelled with propidium iodide (PI) and analysed by flow cytometry by a FACSCalibur flow cytometer (Becton Dickinson, North Ryde, NSW, Australia). DNA damaging agent etopoxide was used as positive control for apoptosis. Results are the mean of three experiments repeated three times.

    Article Snippet: Indomethacin and thapsigargin (Sigma-Aldrich, St. Louis, MO, USA) were used at a concentration of 500 nM, etopoxide (Sigma-Aldrich, St. Louis, MO, USA) was used at 25 μM.

    Techniques: Flow Cytometry, Cytometry, Positive Control

    Phosphorylation level of the JNK protein in the response to thapsigargin (TG) diclofenac (DIC) and indomethacin (IND) in Huh7 cells . Western blot analysis of cell extracts obtained from Huh7 cells exposed to TG, DIC or to IND for the times indicated. Etopoxide was used as positive control for the JNK protein phosphorylation. The blots were probed with antibodies against the unphosphorylated (JNK) or phosphorylated (pJNK) JNK proteins.

    Journal: The Open Biochemistry Journal

    Article Title: In the Huh7 Hepatoma Cells Diclofenac and Indomethacin Activate Differently the Unfolded Protein Response and Induce ER Stress Apoptosis

    doi: 10.2174/1874091X01105010045

    Figure Lengend Snippet: Phosphorylation level of the JNK protein in the response to thapsigargin (TG) diclofenac (DIC) and indomethacin (IND) in Huh7 cells . Western blot analysis of cell extracts obtained from Huh7 cells exposed to TG, DIC or to IND for the times indicated. Etopoxide was used as positive control for the JNK protein phosphorylation. The blots were probed with antibodies against the unphosphorylated (JNK) or phosphorylated (pJNK) JNK proteins.

    Article Snippet: Indomethacin and thapsigargin (Sigma-Aldrich, St. Louis, MO, USA) were used at a concentration of 500 nM, etopoxide (Sigma-Aldrich, St. Louis, MO, USA) was used at 25 μM.

    Techniques: Western Blot, Positive Control

    Precise quantification of individual treatment effects and monitoring of distinct clinical stages. a Imaging visualizes treatment-induced cell loss and regrowth; 10 5 ALL-50 cells/mouse were injected into 10 mice which received a single intraperitoneal dose of Etoposid (VP-16; 50 mg/kg) in week 6 after tumor cell injection, except the control mouse which was treated with PBS. Animals were imaged before treatment (pre-treatment) and 4 and 11 days after treatment; shown is one representative mouse; all mice are shown in Supplemental Figure 11; b, c Imaging visualizes different sensitivities of individual samples towards treatment; ALL-199 (1×10 4 cells/mouse) or ALL-4S (5×10 4 cells/mouse) were injected into 16 mice; mice were randomized in week 4 into one control (n = 4) and two experimental groups (n = 6 each). Control mice received buffer injection, while the other groups were treated once intraperitoneally with either Etoposid (VP-16; 50 mg/kg) or cyclophosphamide (Cyclo; 150 mg/kg) as indicated. Mice were imaged directly before and 4 days after treatment; shown are 3 representative mice of each treatment group before and 4 days after treatment ( b ); shown is the result of quantification of the images; each line represents a single mouse ( c ); * P

    Journal: PLoS ONE

    Article Title: In Vivo Imaging Enables High Resolution Preclinical Trials on Patients' Leukemia Cells Growing in Mice

    doi: 10.1371/journal.pone.0052798

    Figure Lengend Snippet: Precise quantification of individual treatment effects and monitoring of distinct clinical stages. a Imaging visualizes treatment-induced cell loss and regrowth; 10 5 ALL-50 cells/mouse were injected into 10 mice which received a single intraperitoneal dose of Etoposid (VP-16; 50 mg/kg) in week 6 after tumor cell injection, except the control mouse which was treated with PBS. Animals were imaged before treatment (pre-treatment) and 4 and 11 days after treatment; shown is one representative mouse; all mice are shown in Supplemental Figure 11; b, c Imaging visualizes different sensitivities of individual samples towards treatment; ALL-199 (1×10 4 cells/mouse) or ALL-4S (5×10 4 cells/mouse) were injected into 16 mice; mice were randomized in week 4 into one control (n = 4) and two experimental groups (n = 6 each). Control mice received buffer injection, while the other groups were treated once intraperitoneally with either Etoposid (VP-16; 50 mg/kg) or cyclophosphamide (Cyclo; 150 mg/kg) as indicated. Mice were imaged directly before and 4 days after treatment; shown are 3 representative mice of each treatment group before and 4 days after treatment ( b ); shown is the result of quantification of the images; each line represents a single mouse ( c ); * P

    Article Snippet: Preclinical in vivo Treatment Trials Control animals received physiological salt solution intraperitoneally; treatment group mice were injected i.p. with a single dose of either Etoposid (VP-16; 50 mg/kg; Sigma, Hamburg, Germany) or Cyclophosphamide (Cyclo; 150 mg/kg; Baxter, Unterschleissheim, Germany) diluted in 0.9% NaCl.

    Techniques: Imaging, Injection, Mouse Assay