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Becton Dickinson hamster anti human bcl 2
Loss of BAX and BAK or gain of <t>BCL-2</t> blocks cell death in nude mice. Transformed BMK cells expressing red fluorescent protein (RFP) were injected subcutaneously into nude mice. Mice were ear-tagged and individual mice were monitored over time using a whole-body fluorescence imaging system to follow injected BMK cells. One representative animal of each cell line is shown. Note the progressive loss of RFP signal in the control transformed BMK cells (W2.3.1–5), which was largely prevented by the loss of BAX and BAK (D3.zeo-2) or the gain of BCL-2 (W2.Bcl2–3).
Hamster Anti Human Bcl 2, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 85/100, based on 192 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Hypoxia and defective apoptosis drive genomic instability and tumorigenesis"

Article Title: Hypoxia and defective apoptosis drive genomic instability and tumorigenesis

Journal: Genes & Development

doi: 10.1101/gad.1204904

Loss of BAX and BAK or gain of BCL-2 blocks cell death in nude mice. Transformed BMK cells expressing red fluorescent protein (RFP) were injected subcutaneously into nude mice. Mice were ear-tagged and individual mice were monitored over time using a whole-body fluorescence imaging system to follow injected BMK cells. One representative animal of each cell line is shown. Note the progressive loss of RFP signal in the control transformed BMK cells (W2.3.1–5), which was largely prevented by the loss of BAX and BAK (D3.zeo-2) or the gain of BCL-2 (W2.Bcl2–3).
Figure Legend Snippet: Loss of BAX and BAK or gain of BCL-2 blocks cell death in nude mice. Transformed BMK cells expressing red fluorescent protein (RFP) were injected subcutaneously into nude mice. Mice were ear-tagged and individual mice were monitored over time using a whole-body fluorescence imaging system to follow injected BMK cells. One representative animal of each cell line is shown. Note the progressive loss of RFP signal in the control transformed BMK cells (W2.3.1–5), which was largely prevented by the loss of BAX and BAK (D3.zeo-2) or the gain of BCL-2 (W2.Bcl2–3).

Techniques Used: Mouse Assay, Transformation Assay, Expressing, Injection, Fluorescence, Imaging

Formation of tumor giant cells in tumors defective for apoptosis. ( A ) Hematoxylin and eosin-stained sections reveal numerous tumor giant cells in tumors from transformed BMK cells expressing BCL-2 or E1B 19K. Typical sections of carcinomas as described in the text are shown at a magnification of 200× and highlight areas enriched for tumor giant cells in tumors from animals injected with transformed BMK cells expressing BCL-2 or E1B 19K, with insets of grossly polyploid cells in mitosis (at 1000× magnification). Several tumor giant cells in each image are indicated by white arrows. Note the absence of tumor giant cells in tumors formed by the W2.3.1–5 cells. A typical section of a tumor area enriched for tumor giant cells was also immunostained for adenovirus E1A to demonstrate that the tumor giant cells are derived from the input-transformed BMK cells. Note the numerous tumor giant cells that stain brown in the E1A immunohistochemistry, including several examples indicated by arrows. A tumor section (20 μm) enriched for tumor giant cells (arrows) was stained with YOYO-1 to reveal DNA content as described in the text. This image is shown at 630×. ( B ) Aberrant metaphases and polyploid cells accumulate during tumor progression. Tumor sections were developed by immunohistochemistry using antibodies specific for phospho-histone H3 and are shown at 200×. Black arrows indicate aberrant polyploid mitotic arrays, and mitotic arrays presented at 600× in the insets are boxed. Top panels represent images of sections from mature tumors. Phosphohistone H3 immunohistochemistry of sections of transformed BMK cell masses excised from mice on days 2 and 9 after injection are shown in the bottom two rows (200×). Insets in the phospho-histone H3 panels were photographed at 600×, and areas present in the insets are boxed. Grossly aberrant mitotic arrays stained for phospho-histone H3 that are evident in the W2.Bcl2–3 and D3.zeo-2, but not W2.3.1–5, cells on day 9 are indicated in the insets by white arrows. Necrotic centers are indicated (N).
Figure Legend Snippet: Formation of tumor giant cells in tumors defective for apoptosis. ( A ) Hematoxylin and eosin-stained sections reveal numerous tumor giant cells in tumors from transformed BMK cells expressing BCL-2 or E1B 19K. Typical sections of carcinomas as described in the text are shown at a magnification of 200× and highlight areas enriched for tumor giant cells in tumors from animals injected with transformed BMK cells expressing BCL-2 or E1B 19K, with insets of grossly polyploid cells in mitosis (at 1000× magnification). Several tumor giant cells in each image are indicated by white arrows. Note the absence of tumor giant cells in tumors formed by the W2.3.1–5 cells. A typical section of a tumor area enriched for tumor giant cells was also immunostained for adenovirus E1A to demonstrate that the tumor giant cells are derived from the input-transformed BMK cells. Note the numerous tumor giant cells that stain brown in the E1A immunohistochemistry, including several examples indicated by arrows. A tumor section (20 μm) enriched for tumor giant cells (arrows) was stained with YOYO-1 to reveal DNA content as described in the text. This image is shown at 630×. ( B ) Aberrant metaphases and polyploid cells accumulate during tumor progression. Tumor sections were developed by immunohistochemistry using antibodies specific for phospho-histone H3 and are shown at 200×. Black arrows indicate aberrant polyploid mitotic arrays, and mitotic arrays presented at 600× in the insets are boxed. Top panels represent images of sections from mature tumors. Phosphohistone H3 immunohistochemistry of sections of transformed BMK cell masses excised from mice on days 2 and 9 after injection are shown in the bottom two rows (200×). Insets in the phospho-histone H3 panels were photographed at 600×, and areas present in the insets are boxed. Grossly aberrant mitotic arrays stained for phospho-histone H3 that are evident in the W2.Bcl2–3 and D3.zeo-2, but not W2.3.1–5, cells on day 9 are indicated in the insets by white arrows. Necrotic centers are indicated (N).

Techniques Used: Staining, Transformation Assay, Expressing, Injection, Derivative Assay, Immunohistochemistry, Mouse Assay

Antiapoptotic BCL-2 family proteins block apoptosis and promote tumor formation. ( A ) Generation of stable cell lines. Cell extracts made from stable BMK cells that express both BAX and BAK (W2), or that are deficient for BAX and BAK (D3), were subjected to Western blotting with antibodies specific for BCL-2 ( left top panel) or E1B 19K ( right top panel). Note the similar expression levels of each exogenous protein in three independent clones (depicted numerically) and undetectable levels of each exogenous protein in the vector-only control cell lines (W2.3.1–2,5,6 or D3.zeo-1,2,3). Blots were then reprobed with an antibody to actin to verify nearly equivalent levels of protein in all lanes, shown below the BCL-2 and E1B 19K panels. ( B ) BCL-2 and E1B 19K block apoptosis in response to staurosporine. Stable BMK cell lines expressing BCL-2, E1B 19K, and controls were treated with media alone (open bars) or media containing 0.4 μM staurosporine (filled bars) for 24 h, and the viable cell number was determined by trypan blue exclusion. Results are presented as the percent of viable cells in each condition, which in each case represents the average of three independent plates. ( C ) BCL-2 and E1B 19K antagonize BAX and BAK to promote tumor formation. Three independent stable BMK cell lines (circles, squares, and diamonds) expressing BCL-2 (green symbols), E1B 19K (blue symbols), or controls (red symbols) were injected subcutaneously into nude mice, and tumor formation was monitored over time. Each point represents the average tumor volume for five injected animals. W2 cells, which express both BAX and BAK, are shown in the left panel. D3 cells, which are deficient for both BAX and BAK, are shown in the right panel. Note that BCL-2 or E1B 19K expression caused a profound acceleration of tumor formation in the W2 cells, whereas the kinetics of tumor formation in the D3 cells, which are deficient for both BAX and BAK, were unchanged by BCL-2 or E1B 19K expression.
Figure Legend Snippet: Antiapoptotic BCL-2 family proteins block apoptosis and promote tumor formation. ( A ) Generation of stable cell lines. Cell extracts made from stable BMK cells that express both BAX and BAK (W2), or that are deficient for BAX and BAK (D3), were subjected to Western blotting with antibodies specific for BCL-2 ( left top panel) or E1B 19K ( right top panel). Note the similar expression levels of each exogenous protein in three independent clones (depicted numerically) and undetectable levels of each exogenous protein in the vector-only control cell lines (W2.3.1–2,5,6 or D3.zeo-1,2,3). Blots were then reprobed with an antibody to actin to verify nearly equivalent levels of protein in all lanes, shown below the BCL-2 and E1B 19K panels. ( B ) BCL-2 and E1B 19K block apoptosis in response to staurosporine. Stable BMK cell lines expressing BCL-2, E1B 19K, and controls were treated with media alone (open bars) or media containing 0.4 μM staurosporine (filled bars) for 24 h, and the viable cell number was determined by trypan blue exclusion. Results are presented as the percent of viable cells in each condition, which in each case represents the average of three independent plates. ( C ) BCL-2 and E1B 19K antagonize BAX and BAK to promote tumor formation. Three independent stable BMK cell lines (circles, squares, and diamonds) expressing BCL-2 (green symbols), E1B 19K (blue symbols), or controls (red symbols) were injected subcutaneously into nude mice, and tumor formation was monitored over time. Each point represents the average tumor volume for five injected animals. W2 cells, which express both BAX and BAK, are shown in the left panel. D3 cells, which are deficient for both BAX and BAK, are shown in the right panel. Note that BCL-2 or E1B 19K expression caused a profound acceleration of tumor formation in the W2 cells, whereas the kinetics of tumor formation in the D3 cells, which are deficient for both BAX and BAK, were unchanged by BCL-2 or E1B 19K expression.

Techniques Used: Blocking Assay, Stable Transfection, Western Blot, Expressing, Clone Assay, Plasmid Preparation, Injection, Mouse Assay

Defective apoptosis confers resistance to ischemia-induced cell death and allows for the accumulation of polyploid cells in response to ischemia in vitro. ( A ) Gain of BCL-2 or loss of BAX and BAK blocks ischemia-induced cell death. Transformed BMK cell lines were subjected to in vitro ischemic culture conditions and viabilities were determined on the indicated days. Results of triplicates are plotted for each time point. ( B ) PUMA is induced by ischemia in vitro. Cell extracts made from cells cultured for 24 h in normal control conditions (C), or in ischemic culture conditions (I), were immunoblotted for PUMA, BIM, and actin. Note the induction of PUMA, but not BIM, in ischemic culture conditions. ( C,D ) Gain of BCL-2 or loss of BAX and BAK allows for accumulation of polyploid cells in response to ischemia. Transformed BMK cell lines were subjected to ischemic culture conditions for 2 d and then returned to normal culture conditions in complete media for 3 d. ( C ) Cells were collected prior to ischemia, following 2 d of ischemia, and after ischemia with an additional 3 d of recovery on return to normal culture conditions and analyzed by flow cytometry. Insets illustrate typical nuclear morphology in each condition visualized by DAPI staining of cells on coverslips. Arrows indicate typical multi-/giant-nuclei in W2.Bcl2–3 and D3.zeo-2 cells, and the arrowhead indicates a W2.3.1–5 cell dying in hypoxia. ( D ) Representative polyploid cells stained with DAPI were also imaged using confocal laser scanning microscopy. In each panel, a nucleus of normal size cell is included in the picture for comparison. Ratios of the total DNA content of the polyploid cells to the total DNA content of the control cells are included in the insets for each panel.
Figure Legend Snippet: Defective apoptosis confers resistance to ischemia-induced cell death and allows for the accumulation of polyploid cells in response to ischemia in vitro. ( A ) Gain of BCL-2 or loss of BAX and BAK blocks ischemia-induced cell death. Transformed BMK cell lines were subjected to in vitro ischemic culture conditions and viabilities were determined on the indicated days. Results of triplicates are plotted for each time point. ( B ) PUMA is induced by ischemia in vitro. Cell extracts made from cells cultured for 24 h in normal control conditions (C), or in ischemic culture conditions (I), were immunoblotted for PUMA, BIM, and actin. Note the induction of PUMA, but not BIM, in ischemic culture conditions. ( C,D ) Gain of BCL-2 or loss of BAX and BAK allows for accumulation of polyploid cells in response to ischemia. Transformed BMK cell lines were subjected to ischemic culture conditions for 2 d and then returned to normal culture conditions in complete media for 3 d. ( C ) Cells were collected prior to ischemia, following 2 d of ischemia, and after ischemia with an additional 3 d of recovery on return to normal culture conditions and analyzed by flow cytometry. Insets illustrate typical nuclear morphology in each condition visualized by DAPI staining of cells on coverslips. Arrows indicate typical multi-/giant-nuclei in W2.Bcl2–3 and D3.zeo-2 cells, and the arrowhead indicates a W2.3.1–5 cell dying in hypoxia. ( D ) Representative polyploid cells stained with DAPI were also imaged using confocal laser scanning microscopy. In each panel, a nucleus of normal size cell is included in the picture for comparison. Ratios of the total DNA content of the polyploid cells to the total DNA content of the control cells are included in the insets for each panel.

Techniques Used: In Vitro, Transformation Assay, Cell Culture, Flow Cytometry, Cytometry, Staining, Confocal Laser Scanning Microscopy

2) Product Images from "Evaluation of apoptogenic adenovirus type 5 oncolytic vectors in a Syrian hamster head and neck cancer model"

Article Title: Evaluation of apoptogenic adenovirus type 5 oncolytic vectors in a Syrian hamster head and neck cancer model

Journal: Cancer gene therapy

doi: 10.1038/cgt.2014.22

Apoptotic activity of HAdV5 mutants in hamster cells (A). Effect of HAdV5 mutants on caspase activation. HPT11 and 12 cells were infected with indicated mutants at 400 PFU/cell and proteolytic processing of pro-caspase 3, EGFR and PARP were determined by western blot analysis 24 hr after infection. (B). HPT11 and 12 cells were infected 400 PFU/cell and the annexin V-staining patterns were determined by FACS analysis. A representative analysis is shown in B. (C). Averages of annexin V-positive cells were shown as an indication of apoptotic cell death. The experiment was repeated at least three times independently and the data are means ±SD. Statistical differences were assessed using one-way ANOVA followed by a Newman-Keuls Multiple Comparision Test, where appropriate, using GraphPad Prism 5 software. P- Value was determined by comparing mock, dl 312 and Ad5 wt vs lp 11w and mock, dl 312 and Ad5 wt vs lp 11w/Δ55K in both HPT11 and HPT12 cell lines. Statistical significance was accepted at a value of * P
Figure Legend Snippet: Apoptotic activity of HAdV5 mutants in hamster cells (A). Effect of HAdV5 mutants on caspase activation. HPT11 and 12 cells were infected with indicated mutants at 400 PFU/cell and proteolytic processing of pro-caspase 3, EGFR and PARP were determined by western blot analysis 24 hr after infection. (B). HPT11 and 12 cells were infected 400 PFU/cell and the annexin V-staining patterns were determined by FACS analysis. A representative analysis is shown in B. (C). Averages of annexin V-positive cells were shown as an indication of apoptotic cell death. The experiment was repeated at least three times independently and the data are means ±SD. Statistical differences were assessed using one-way ANOVA followed by a Newman-Keuls Multiple Comparision Test, where appropriate, using GraphPad Prism 5 software. P- Value was determined by comparing mock, dl 312 and Ad5 wt vs lp 11w and mock, dl 312 and Ad5 wt vs lp 11w/Δ55K in both HPT11 and HPT12 cell lines. Statistical significance was accepted at a value of * P

Techniques Used: Activity Assay, Activation Assay, Infection, Western Blot, Staining, FACS, Software

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