bcl-2 Santa Cruz Biotechnology Search Results


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  • 99
    Santa Cruz Biotechnology bcl 2
    The schematic diagram illustrates that oral treatment with the herbal formula B307 alleviates cardiotoxicity in doxorubicin-treated mice via suppressing hypoxia, oxidative stress, inflammation, and apoptosis in the heart tissue. Abbreviations: eNOS, endothelial nitric oxide synthase; SOD2, superoxide dismutase 2; ROS, reactive oxygen species; 3-NT, neurotrophin-3; 4-HNE, 4-hydroxynonenal; NF-κB, nuclear factor-κB; TNF-α, tumor necrosis factor alpha; er, endoplasmic reticulum; <t>Bcl-2,</t> B-cell lymphoma 2; Bax, Bcl-2-associated X protein; Cyto-C, cytochrome C.
    Bcl 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 99/100, based on 11549 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology ser 70 phospho bcl 2
    The schematic diagram illustrates that oral treatment with the herbal formula B307 alleviates cardiotoxicity in doxorubicin-treated mice via suppressing hypoxia, oxidative stress, inflammation, and apoptosis in the heart tissue. Abbreviations: eNOS, endothelial nitric oxide synthase; SOD2, superoxide dismutase 2; ROS, reactive oxygen species; 3-NT, neurotrophin-3; 4-HNE, 4-hydroxynonenal; NF-κB, nuclear factor-κB; TNF-α, tumor necrosis factor alpha; er, endoplasmic reticulum; <t>Bcl-2,</t> B-cell lymphoma 2; Bax, Bcl-2-associated X protein; Cyto-C, cytochrome C.
    Ser 70 Phospho Bcl 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 75/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology antibodies against bcl 2
    Effect of co-treatment of breast cancer cell lines with 4-hydroxytamoxifen and erucin on PARP cleavage, levels of <t>Bcl-2,</t> Bax, survivin and ADRP. T47D ( a ) and MCF-7 ( b ) cells were treated with 5 μM erucin (ERN) and/or 0.5 μM 4-hydroxytamoxifen (4-OH-T). BT-474 ( c ) cells were treated with 5 μM erucin (ERN) and/or 1 μM 4-hydroxytamoxifen (4-OH-T). Blots were stripped and reprobed with anti-β-actin antibody to ensure equal protein loading. Results are plotted as mean ± SE from 3 independent experiments, *significantly different compared with single agent-treated samples or **significantly different compared with one of the single agent-treated samples by one-way ANOVA followed by Bonferroni’s multiple comparison test. Data for PARP refer to the faster migrating band marked as * and are given relative to samples treated with ERN alone. Blots shown are representative of at least three independent experiments
    Antibodies Against Bcl 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 99/100, based on 496 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology b cell lymphoma 2
    Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL), Bax, B-cell lymphoma 2-associated X protein (Bax), and B-cell <t>lymphoma</t> 2 (Bcl-2) staining in the penumbra, and the effects of cilostazol (CZL) after 96 hours in Long-Evans Tokushima Otsuka (LETO) and Otsuka Long-Evans Tokushima Fatty (OLETF) rats. The number of apoptotic TUNEL-positive cells (with darkly stained nuclei or nuclear fragments with a cytoplasmic halo) was reduced by CZL. Strong Bax protein expression was observed both in the penumbra and in areas distant from the acute ischemic lesion, and was suppressed by CZL. Immunolabeling for Bcl-2 near the infarct area was increased by CZL (bar=200 μ m, × 200 magnification). CMC, carboxymethyl cellulose.
    B Cell Lymphoma 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 99/100, based on 680 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Santa Cruz Biotechnology human bcl 2
    Stable expression of human <t>Bcl-2</t> significantly reduces NS1-induced cell death. (A) Representative Bcl-2-stable transfectants of both KLNS and ULNS cells were detected with a Bcl-2 MAb, Bcl-2 100, by immunoblotting with 5 × 10 6 cells in the presence or absence of IPTG. The position of the Bcl-2 protein corresponding to approximately 26 kDa is indicated. (B and C) The kinetics of Bcl-2 inhibition of NS1-induced cell death was measured by trypan blue exclusion for both KLNS (B) and ULNS (C) cells with 5 × 10 5 cells for each cell line. The experiments were performed twice, with no significant difference between results.
    Human Bcl 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 121 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology mab against bcl 2
    Stable expression of human <t>Bcl-2</t> significantly reduces NS1-induced cell death. (A) Representative Bcl-2-stable transfectants of both KLNS and ULNS cells were detected with a Bcl-2 MAb, Bcl-2 100, by immunoblotting with 5 × 10 6 cells in the presence or absence of IPTG. The position of the Bcl-2 protein corresponding to approximately 26 kDa is indicated. (B and C) The kinetics of Bcl-2 inhibition of NS1-induced cell death was measured by trypan blue exclusion for both KLNS (B) and ULNS (C) cells with 5 × 10 5 cells for each cell line. The experiments were performed twice, with no significant difference between results.
    Mab Against Bcl 2, supplied by Santa Cruz Biotechnology, 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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    Santa Cruz Biotechnology b cell lymphoma 2 bcl 2
    The schematic diagram illustrates the possible neuroprotective pathways in the brain under oral B401 treatment. Oral B401 treatment may improve neuroprotection in R6/2 mice via enhancing anti-oxidative stress (marked by SOD2) and anti-apoptosis (marked by <t>Bcl-2),</t> while suppressing mutant huntingtin aggregation, ROS production, ER stress-related apoptosis (marked by calpain), and mitochondrial dysfunction-related apoptosis (marked by Bax and caspase 3) in the brain. Abbreviations: Bax, Bcl-2-associated X protein; Bcl-2, B-cell lymphoma 2; ER, endoplasmic reticulum; mHtt, mutant huntingtin; ROS, reactive oxygen species; SOD2, superoxide dismutase 2.
    B Cell Lymphoma 2 Bcl 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 91 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology rabbit polyclonal antibody against bcl 2
    The schematic diagram illustrates the possible neuroprotective pathways in the brain under oral B401 treatment. Oral B401 treatment may improve neuroprotection in R6/2 mice via enhancing anti-oxidative stress (marked by SOD2) and anti-apoptosis (marked by <t>Bcl-2),</t> while suppressing mutant huntingtin aggregation, ROS production, ER stress-related apoptosis (marked by calpain), and mitochondrial dysfunction-related apoptosis (marked by Bax and caspase 3) in the brain. Abbreviations: Bax, Bcl-2-associated X protein; Bcl-2, B-cell lymphoma 2; ER, endoplasmic reticulum; mHtt, mutant huntingtin; ROS, reactive oxygen species; SOD2, superoxide dismutase 2.
    Rabbit Polyclonal Antibody Against Bcl 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology monoclonal antibodies against bcl 2
    Increase in chemotherapeutic drug-induced apoptosis by miR-218. SGC7901 cells were transfected with miR-218 mimic or the negative control and incubated 48 h with (A) ADM or (B) L-OHP. Proportions of apoptotic cells under the different conditions were determined using flow cytometry. Expression levels of <t>Bcl-2</t> (C) and Bax (D) in SGC7901 cells were examined 48 h after transfection. Results are shown from three independent experiments. * P
    Monoclonal Antibodies Against Bcl 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology primary antibodies against bcl 2
    Effect of PD on the expression of apoptosis-related proteins. (A) A549 human lung cancer cells were treated with increasing concentrations of PD for 24 and 48 h. Expression levels of <t>Bcl-2</t> and Bax were detected by western blot analysis and β-actin was used as a control. (B and C) Bcl-2 and (D and E) Bax protein bands were quantitated by densitometry, and the data are presented as the mean ± SD from three experiments. * P
    Primary Antibodies Against Bcl 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology polyclonal antibodies against bcl 2
    Effect of PD on the expression of apoptosis-related proteins. (A) A549 human lung cancer cells were treated with increasing concentrations of PD for 24 and 48 h. Expression levels of <t>Bcl-2</t> and Bax were detected by western blot analysis and β-actin was used as a control. (B and C) Bcl-2 and (D and E) Bax protein bands were quantitated by densitometry, and the data are presented as the mean ± SD from three experiments. * P
    Polyclonal Antibodies Against Bcl 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology mouse bcl 2
    Western blot analysis. Serine phosphorylation and <t>Bcl-2</t> expression in spleen cells cultured in the presence and absence of T-CHO. (a) Serine phosphorylation; (b) Bcl-2 expression after 24 hr in culture and 96 hr in culture. Lanes 1 and 5, spleen cells from pre-exposed mice, cultured in the presence of T-CHO; lanes 2 and 4, cells from T-CHO pre-exposed mice, cultured in the absence T-CHO; lanes 3 and 7, cells from naive mice cultured in the presence of T-CHO and lanes 4 and 8, cells from naive mice cultured in the absence of T-CHO.
    Mouse Bcl 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology b cell leukemia lymphoma 2 bcl 2
    Western blot analysis. Serine phosphorylation and <t>Bcl-2</t> expression in spleen cells cultured in the presence and absence of T-CHO. (a) Serine phosphorylation; (b) Bcl-2 expression after 24 hr in culture and 96 hr in culture. Lanes 1 and 5, spleen cells from pre-exposed mice, cultured in the presence of T-CHO; lanes 2 and 4, cells from T-CHO pre-exposed mice, cultured in the absence T-CHO; lanes 3 and 7, cells from naive mice cultured in the presence of T-CHO and lanes 4 and 8, cells from naive mice cultured in the absence of T-CHO.
    B Cell Leukemia Lymphoma 2 Bcl 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology mouse anti bcl 2
    Western blotting studies. ( A ). Western blotting analysis of pAKT, tAKT, PCNA, MMP9, <t>Bcl-2,</t> Bax, p53, cleaved caspase-3 with β-actin as an internal reference. ( B ). Quantification of the Western blotting results. Data was presented as the mean ± SD. *p
    Mouse Anti Bcl 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 99/100, based on 283 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology anti human bcl 2
    Analysis of B lymphocyte populations in wild-type vs. transgenic mice. Four-color flow-cytometry analysis was performed to determine the phenotype of B lymphocytes. Gating was performed on the B220 + population (R1 and R2, Left ) in the case of CD23/CD21 and IgM/IgD analyses or on the lymphocyte population for B220/CD5 and isotype control analyses. Splenocytes were analyzed from representative 11-month-old wild-type, <t>Bcl-2,</t> TRAF2DN, and TRAF2DN/Bcl-2 double-transgenic mice at a premalignant stage and splenocytes or blood lymphocytes of a representative TRAF2DN/Bcl-2 mouse (12 months old) with acute disease.
    Anti Human Bcl 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 81/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology anti b cell leukaemia lymphoma 2
    Analysis of B lymphocyte populations in wild-type vs. transgenic mice. Four-color flow-cytometry analysis was performed to determine the phenotype of B lymphocytes. Gating was performed on the B220 + population (R1 and R2, Left ) in the case of CD23/CD21 and IgM/IgD analyses or on the lymphocyte population for B220/CD5 and isotype control analyses. Splenocytes were analyzed from representative 11-month-old wild-type, <t>Bcl-2,</t> TRAF2DN, and TRAF2DN/Bcl-2 double-transgenic mice at a premalignant stage and splenocytes or blood lymphocytes of a representative TRAF2DN/Bcl-2 mouse (12 months old) with acute disease.
    Anti B Cell Leukaemia Lymphoma 2, supplied by Santa Cruz Biotechnology, 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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    Santa Cruz Biotechnology b cell leukemia lymphoma 2
    SAR131675 attenuates apoptosis and oxidative stress in the kidneys in  db/db  mice. Glomerulosclerosis and tubulointerstitial fibrosis were determined at 20 weeks in  db/m  and  db/db  mice treated with or without SAR131675.  a  Representative images of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining in the glomeruli and tubules and quantitative data.  b  Representative western blots for B cell leukemia/lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and β-actin in the kidneys and quantitative data.  c  Twenty-four hour urinary 8-hydroxy-deoxyguanosine (8-OH-dG) and isoprostane levels. * P
    B Cell Leukemia Lymphoma 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 89/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology rabbit anti b cell lymphoma 2
    SAR131675 attenuates apoptosis and oxidative stress in the kidneys in  db/db  mice. Glomerulosclerosis and tubulointerstitial fibrosis were determined at 20 weeks in  db/m  and  db/db  mice treated with or without SAR131675.  a  Representative images of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining in the glomeruli and tubules and quantitative data.  b  Representative western blots for B cell leukemia/lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and β-actin in the kidneys and quantitative data.  c  Twenty-four hour urinary 8-hydroxy-deoxyguanosine (8-OH-dG) and isoprostane levels. * P
    Rabbit Anti B Cell Lymphoma 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 87/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology anti mouse human bcl 2
    Peptide-induced Ca 2+ oscillations in primary CLL cells. (A) Immunoblot comparing <t>Bcl-2</t> levels in normal human B lymphocytes and 3 primary CLL samples. (B-D) Representative single-cell Ca 2+ recordings illustrating the Ca 2+ responses observed in CLL cells
    Anti Mouse Human Bcl 2, supplied by Santa Cruz Biotechnology, 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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    Santa Cruz Biotechnology mouse polyclonal antibody against bcl 2
    Peptide-induced Ca 2+ oscillations in primary CLL cells. (A) Immunoblot comparing <t>Bcl-2</t> levels in normal human B lymphocytes and 3 primary CLL samples. (B-D) Representative single-cell Ca 2+ recordings illustrating the Ca 2+ responses observed in CLL cells
    Mouse Polyclonal Antibody Against Bcl 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 80/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology mouse monoclonal antibodies against bcl 2
    M11L specifically binds to Bak but not to other <t>Bcl-2</t> family members by use of the TAP method. (A) TAP method for isolating M11L binding partners. The plasmid (M11L-C-TAP) was transiently transfected into 293T cells by the calcium phosphate method (step 1). The lysate with putative TAP M11L binding partner complexes was incubated with rabbit agarose IgG beads in order to bind the protein A of the TAP cassette. The beads were washed to remove unbound proteins and then incubated with TEV protease (step 2) to release bound complexes. The protein complex from cleaved products was allowed to bind via calmodulin binding protein (CBP) resin, and then the purified complex was eluted with calmodulin elution buffer containing EGTA (step 3). The eluants, containing proteins interacting with M11L, were separated by SDS-PAGE, and the proteins were analyzed (step 4). (B) Bak, but not Bad, Bid, Bax/p21, or Bcl-2, was detected as a binding partner of M11L in the M11L-C-TAP eluted products (lane 3), but not in control eluants of C-TAP alone (lane 2), following lysis with CHAPS buffer. Lane 1 confirms endogenous expression levels of the various Bcl-2 family members in 293T cell lysates. (C) Cell lysates of the C-TAP vector alone (lane 1) or the M11L-C-TAP bait (lane 2) were probed with anti-PAP before the TAP procedure to confirm the expression and size of the M11L-TAP fusion protein (34 kDa). (D) Eluants of the C-TAP vector alone (lane 1) or M11L-C-TAP (lane 2) were immunoblotted with anti-M11L, indicating that the M11L bait tagged with CBP (17 kDa) was recovered following TEV cleavage and elution. (E) huBak binds to untagged M11L. Shown are TAP eluants from lysates of cotransfections of pcDNA3-M11L plus N-TAP vector alone (lane 1) or pcDNA3-M11L plus N-TAP-huBak (lane 2) in 293T cells. Lane 3 (M11L and TAP vector) and lane 4 (M11L and N-TAP-huBak) represent corresponding cell lysates, confirming the expression of the appropriate cotransfected plasmids.
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    Santa Cruz Biotechnology fitc conjugated monoclonal antibody against bcl 2
    M11L specifically binds to Bak but not to other <t>Bcl-2</t> family members by use of the TAP method. (A) TAP method for isolating M11L binding partners. The plasmid (M11L-C-TAP) was transiently transfected into 293T cells by the calcium phosphate method (step 1). The lysate with putative TAP M11L binding partner complexes was incubated with rabbit agarose IgG beads in order to bind the protein A of the TAP cassette. The beads were washed to remove unbound proteins and then incubated with TEV protease (step 2) to release bound complexes. The protein complex from cleaved products was allowed to bind via calmodulin binding protein (CBP) resin, and then the purified complex was eluted with calmodulin elution buffer containing EGTA (step 3). The eluants, containing proteins interacting with M11L, were separated by SDS-PAGE, and the proteins were analyzed (step 4). (B) Bak, but not Bad, Bid, Bax/p21, or Bcl-2, was detected as a binding partner of M11L in the M11L-C-TAP eluted products (lane 3), but not in control eluants of C-TAP alone (lane 2), following lysis with CHAPS buffer. Lane 1 confirms endogenous expression levels of the various Bcl-2 family members in 293T cell lysates. (C) Cell lysates of the C-TAP vector alone (lane 1) or the M11L-C-TAP bait (lane 2) were probed with anti-PAP before the TAP procedure to confirm the expression and size of the M11L-TAP fusion protein (34 kDa). (D) Eluants of the C-TAP vector alone (lane 1) or M11L-C-TAP (lane 2) were immunoblotted with anti-M11L, indicating that the M11L bait tagged with CBP (17 kDa) was recovered following TEV cleavage and elution. (E) huBak binds to untagged M11L. Shown are TAP eluants from lysates of cotransfections of pcDNA3-M11L plus N-TAP vector alone (lane 1) or pcDNA3-M11L plus N-TAP-huBak (lane 2) in 293T cells. Lane 3 (M11L and TAP vector) and lane 4 (M11L and N-TAP-huBak) represent corresponding cell lysates, confirming the expression of the appropriate cotransfected plasmids.
    Fitc Conjugated Monoclonal Antibody Against Bcl 2, supplied by Santa Cruz Biotechnology, 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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    Santa Cruz Biotechnology rabbit anti bcl 2
    Temporal changes of autophagy and apoptosis after tGCI. a Immunoblots of the apoptosis-related proteins cleaved Caspase-3 and <t>Bcl-2.</t> Total cell lysates from the hippocampus were used, and β-actin was used as an internal control. b Immunoblots of p53, Bax, and cytochrome c in the cytosolic and mitochondrial fractions. GAPDH was used as a loading control for cytosolic proteins, and COX IV was used as the loading control for mitochondrial proteins. c Immunoblots of the autophagy-related proteins LC3B and Beclin-1. Total cell lysates from the hippocampus were used. d – k Densitometric analysis (mean ± SEM, n = 3 animals per group) of the proteins from ( a , b , c ) normalized to the respective loading controls. * P
    Rabbit Anti Bcl 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 266 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology mab100
    Temporal changes of autophagy and apoptosis after tGCI. a Immunoblots of the apoptosis-related proteins cleaved Caspase-3 and <t>Bcl-2.</t> Total cell lysates from the hippocampus were used, and β-actin was used as an internal control. b Immunoblots of p53, Bax, and cytochrome c in the cytosolic and mitochondrial fractions. GAPDH was used as a loading control for cytosolic proteins, and COX IV was used as the loading control for mitochondrial proteins. c Immunoblots of the autophagy-related proteins LC3B and Beclin-1. Total cell lysates from the hippocampus were used. d – k Densitometric analysis (mean ± SEM, n = 3 animals per group) of the proteins from ( a , b , c ) normalized to the respective loading controls. * P
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    Santa Cruz Biotechnology goat anti bcl 2
    Effect of EGCG on protein expression of apoptotic markers Bax and <t>Bcl-2</t> in kidney of control and fluoride-treated rats. Protein expression was determined by Western blot analyses. Values are expressed as mean ± SD for measurements from six rats in each group. Statistical significance was determined by one way ANOVA followed by post hoc t -test. Bars with different superscript letters (a–c) differ significantly at P
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    Santa Cruz Biotechnology polyclonal anti human b cell lymphoma 2
    Effect of EGCG on protein expression of apoptotic markers Bax and <t>Bcl-2</t> in kidney of control and fluoride-treated rats. Protein expression was determined by Western blot analyses. Values are expressed as mean ± SD for measurements from six rats in each group. Statistical significance was determined by one way ANOVA followed by post hoc t -test. Bars with different superscript letters (a–c) differ significantly at P
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    Santa Cruz Biotechnology polyclonal mouse anti human bcl 2
    Effect of EGCG on protein expression of apoptotic markers Bax and <t>Bcl-2</t> in kidney of control and fluoride-treated rats. Protein expression was determined by Western blot analyses. Values are expressed as mean ± SD for measurements from six rats in each group. Statistical significance was determined by one way ANOVA followed by post hoc t -test. Bars with different superscript letters (a–c) differ significantly at P
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    Santa Cruz Biotechnology endogenous mouse bcl 2
    <t>RCAS-Bcl-2</t> decreases the apoptotic index in anaplastic oligodendroglioma
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    Santa Cruz Biotechnology rabbit anti human bcl 2
    Proposed model of targeting therapy and signaling pathway involved in CGOML-induced apoptosis of MCF-7 cells: Nude mice bearing breast cancer (MCF-7) received intravenous injections of GML (35 μg of Gemcitabine/g) at day 1, 5 and then these mice also received intravenous injections of OML (5 μg of oxaliplatin/g) at day 3, 7; magnetic field (5000 GS) was applied to the tumor surface for 30 min after every injection The magnetic properties of GML or OML particles guide the gemcitabine or oxaliplatin to the tumor area of nude mice under an external magnetic field (Nd 2 Fe 12 B magnet tablets). GML or OML has excellent half-life periods and can be gradually biodegraded in MCF-7 cells, lead to gemcitabine of GML (or oxaliplatin of OML) slowly released in target cells. In MCF-7 cells, OML inhibited the replication and transcription of DNA by forming inter-strand and intra-strand platinum-DNA adducts (Pt-GG and Pt-AG), while GML prevented the synthesis and repair of DNA by inhibiting activation of ribonucleotide reductase, and stopped the the synthesis of DNA and broken strands of DNA by replacing the cytidine of DNA strands with dFdCTP. It leads to the phenomenon of DNA ladder, increase of BAX and decrease of <t>Bcl-2</t> and Survivin, eventual activation of caspase-9 and caspase-3 cause cell apoptosis in MCF-7 cells, resulting in MCF-7 cell death.
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    Santa Cruz Biotechnology b cell cll lymphoma 2
    Proposed model of targeting therapy and signaling pathway involved in CGOML-induced apoptosis of MCF-7 cells: Nude mice bearing breast cancer (MCF-7) received intravenous injections of GML (35 μg of Gemcitabine/g) at day 1, 5 and then these mice also received intravenous injections of OML (5 μg of oxaliplatin/g) at day 3, 7; magnetic field (5000 GS) was applied to the tumor surface for 30 min after every injection The magnetic properties of GML or OML particles guide the gemcitabine or oxaliplatin to the tumor area of nude mice under an external magnetic field (Nd 2 Fe 12 B magnet tablets). GML or OML has excellent half-life periods and can be gradually biodegraded in MCF-7 cells, lead to gemcitabine of GML (or oxaliplatin of OML) slowly released in target cells. In MCF-7 cells, OML inhibited the replication and transcription of DNA by forming inter-strand and intra-strand platinum-DNA adducts (Pt-GG and Pt-AG), while GML prevented the synthesis and repair of DNA by inhibiting activation of ribonucleotide reductase, and stopped the the synthesis of DNA and broken strands of DNA by replacing the cytidine of DNA strands with dFdCTP. It leads to the phenomenon of DNA ladder, increase of BAX and decrease of <t>Bcl-2</t> and Survivin, eventual activation of caspase-9 and caspase-3 cause cell apoptosis in MCF-7 cells, resulting in MCF-7 cell death.
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    Santa Cruz Biotechnology bcl 2 associated x protein
    Western blot analysis of Bax, <t>Bcl-2</t> and Bcl-XL protein expression levels in resveratrol-treated HeLa cells. (A) Cells were exposed to 0, 5, 10, 20 and 40 µmol/l of resveratrol for 48 h. (B) Cells were treated with 20 µmol/l resveratrol for 0, 12, 24, 36 and 48 h. All cells were lysed, equal quantities of protein were separated using 12% SDS-PAGE and protein expression was detected by western blotting. Bcl-2, B-cell lymphoma 2; Bax, B-cell lymphoma 2-associated X protein; Bcl-2 XL, B-cell lymphoma 2-extra large.
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    Santa Cruz Biotechnology polyclonal rabbit bcl 2
    Western blot analysis of Bax, <t>Bcl-2</t> and Bcl-XL protein expression levels in resveratrol-treated HeLa cells. (A) Cells were exposed to 0, 5, 10, 20 and 40 µmol/l of resveratrol for 48 h. (B) Cells were treated with 20 µmol/l resveratrol for 0, 12, 24, 36 and 48 h. All cells were lysed, equal quantities of protein were separated using 12% SDS-PAGE and protein expression was detected by western blotting. Bcl-2, B-cell lymphoma 2; Bax, B-cell lymphoma 2-associated X protein; Bcl-2 XL, B-cell lymphoma 2-extra large.
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    Image Search Results


    The schematic diagram illustrates that oral treatment with the herbal formula B307 alleviates cardiotoxicity in doxorubicin-treated mice via suppressing hypoxia, oxidative stress, inflammation, and apoptosis in the heart tissue. Abbreviations: eNOS, endothelial nitric oxide synthase; SOD2, superoxide dismutase 2; ROS, reactive oxygen species; 3-NT, neurotrophin-3; 4-HNE, 4-hydroxynonenal; NF-κB, nuclear factor-κB; TNF-α, tumor necrosis factor alpha; er, endoplasmic reticulum; Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-associated X protein; Cyto-C, cytochrome C.

    Journal: OncoTargets and therapy

    Article Title: Oral treatment with the herbal formula B307 alleviates cardiac toxicity in doxorubicin-treated mice via suppressing oxidative stress, inflammation, and apoptosis

    doi: 10.2147/OTT.S82936

    Figure Lengend Snippet: The schematic diagram illustrates that oral treatment with the herbal formula B307 alleviates cardiotoxicity in doxorubicin-treated mice via suppressing hypoxia, oxidative stress, inflammation, and apoptosis in the heart tissue. Abbreviations: eNOS, endothelial nitric oxide synthase; SOD2, superoxide dismutase 2; ROS, reactive oxygen species; 3-NT, neurotrophin-3; 4-HNE, 4-hydroxynonenal; NF-κB, nuclear factor-κB; TNF-α, tumor necrosis factor alpha; er, endoplasmic reticulum; Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-associated X protein; Cyto-C, cytochrome C.

    Article Snippet: As to the DOX-treated mice, cardiac expression levels of Bcl-2 were visibly enhanced under oral B307 treatment.

    Techniques: Mouse Assay

    Cardiac levels of Bcl-2, a marker of anti-apoptosis, and Bax and Cyto-C, two markers of apoptosis, in mice under sham, B307, DOX, and B307+DOX treatments. Notes: ( A ) IHC staining illustrates that cardiac expression levels of Bcl-2 in the mice were visibly enhanced under oral B307 treatment but were visibly reduced under DOX treatment. As to the DOX-treated mice, cardiac expression levels of Bcl-2 were visibly enhanced under oral B307 treatment. On the other hand, cardiac expression levels of Bax and Cyto-C in the mice were visibly reduced under oral B307 treatment but were visibly enhanced under DOX treatment. As to the DOX-treated mice, cardiac expression levels of Bax and Cyto-C were visibly reduced under oral B307 treatment. ( B ) Western blotting analysis shows the following: (a) expression levels of cardiac Bcl-2, Bax, and Cyto-C under sham, B307, DOX, and B307+DOX treatments and (b) the quantified ratio of Bcl-2/Bax in the heart tissue of the mice was significantly increased under oral B307 treatment but were significantly reduced under DOX treatment. As to the DOX-treated mice, the quantified ratio of Bcl-2/Bax in the heart tissue was significantly increased under oral B307 treatment. On the other hand, quantified Cyto-C levels in the heart tissue of the mice were significantly decreased under oral B307 treatment but were significantly enhanced under DOX treatment. As to the DOX-treated mice, quantified Cyto-C levels in the heart tissue were significantly reduced under oral B307 treatment. The number of mice under sham, B307, DOX, and B307+DOX treatments was six for each group. Values are mean ± SEM (* P

    Journal: OncoTargets and therapy

    Article Title: Oral treatment with the herbal formula B307 alleviates cardiac toxicity in doxorubicin-treated mice via suppressing oxidative stress, inflammation, and apoptosis

    doi: 10.2147/OTT.S82936

    Figure Lengend Snippet: Cardiac levels of Bcl-2, a marker of anti-apoptosis, and Bax and Cyto-C, two markers of apoptosis, in mice under sham, B307, DOX, and B307+DOX treatments. Notes: ( A ) IHC staining illustrates that cardiac expression levels of Bcl-2 in the mice were visibly enhanced under oral B307 treatment but were visibly reduced under DOX treatment. As to the DOX-treated mice, cardiac expression levels of Bcl-2 were visibly enhanced under oral B307 treatment. On the other hand, cardiac expression levels of Bax and Cyto-C in the mice were visibly reduced under oral B307 treatment but were visibly enhanced under DOX treatment. As to the DOX-treated mice, cardiac expression levels of Bax and Cyto-C were visibly reduced under oral B307 treatment. ( B ) Western blotting analysis shows the following: (a) expression levels of cardiac Bcl-2, Bax, and Cyto-C under sham, B307, DOX, and B307+DOX treatments and (b) the quantified ratio of Bcl-2/Bax in the heart tissue of the mice was significantly increased under oral B307 treatment but were significantly reduced under DOX treatment. As to the DOX-treated mice, the quantified ratio of Bcl-2/Bax in the heart tissue was significantly increased under oral B307 treatment. On the other hand, quantified Cyto-C levels in the heart tissue of the mice were significantly decreased under oral B307 treatment but were significantly enhanced under DOX treatment. As to the DOX-treated mice, quantified Cyto-C levels in the heart tissue were significantly reduced under oral B307 treatment. The number of mice under sham, B307, DOX, and B307+DOX treatments was six for each group. Values are mean ± SEM (* P

    Article Snippet: As to the DOX-treated mice, cardiac expression levels of Bcl-2 were visibly enhanced under oral B307 treatment.

    Techniques: Marker, Mouse Assay, Immunohistochemistry, Staining, Expressing, Western Blot

    Glucocorticoid receptor (GR) antagonism downregulates the expression of antiapoptotic Bcl-2 and Bcl-xL proteins. (A) Docetaxel-resistant cells undergo apoptosis upon treatment with RU-486 (3 μM) and docetaxel (30 nM). *** P

    Journal: Endocrine-Related Cancer

    Article Title: Glucocorticoid receptor antagonism reverts docetaxel resistance in human prostate cancer

    doi: 10.1530/ERC-15-0343

    Figure Lengend Snippet: Glucocorticoid receptor (GR) antagonism downregulates the expression of antiapoptotic Bcl-2 and Bcl-xL proteins. (A) Docetaxel-resistant cells undergo apoptosis upon treatment with RU-486 (3 μM) and docetaxel (30 nM). *** P

    Article Snippet: The following antibodies were used: anti-GR (1:500, sc-8992, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-GAPDH (1:10 000, Chemicon, Temecula, CA, USA), anti-β-actin (1:5000, Sigma), anti-Bcl-xL (1:1000, sc-23958, Santa Cruz Biotechnology), and anti-Bcl-2 (1:1000, sc-7382, Santa Cruz Biotechnology).

    Techniques: Expressing

    RNA interference mediated knock-down of Mcl-1 induces cell death in HER2 overexpressing cells . BT474 cells were left untreated or transfected with control siRNA, Bcl-xL siRNA, Bcl-2 siRNA or Mcl-1 siRNA as indicated. (A) Down regulation of the targeted proteins 48 hours after transfection was confirmed by western blot analysis, using tubulin as a loading control. (B) Forty eight hours after transfection, apoptosis was assessed by Apo 2.7 staining and FACS analysis as described in Materials and Methods. Data represented (% of Apo2.7 positive cells) are the means +/- standard errors of three independent experiments. p-values were assessed using a Student's t test. (C) Experiments were performed as described in B, using HER2 overexpressing SKBR3 cells, and ER expressing MCF7 cells, as indicated. Data are mean ± se of the indicated number of independent experiments. p-values were assessed using a Student's t test

    Journal: Molecular Cancer

    Article Title: c-Myc dependent expression of pro-apoptotic Bim renders HER2-overexpressing breast cancer cells dependent on anti-apoptotic Mcl-1

    doi: 10.1186/1476-4598-10-110

    Figure Lengend Snippet: RNA interference mediated knock-down of Mcl-1 induces cell death in HER2 overexpressing cells . BT474 cells were left untreated or transfected with control siRNA, Bcl-xL siRNA, Bcl-2 siRNA or Mcl-1 siRNA as indicated. (A) Down regulation of the targeted proteins 48 hours after transfection was confirmed by western blot analysis, using tubulin as a loading control. (B) Forty eight hours after transfection, apoptosis was assessed by Apo 2.7 staining and FACS analysis as described in Materials and Methods. Data represented (% of Apo2.7 positive cells) are the means +/- standard errors of three independent experiments. p-values were assessed using a Student's t test. (C) Experiments were performed as described in B, using HER2 overexpressing SKBR3 cells, and ER expressing MCF7 cells, as indicated. Data are mean ± se of the indicated number of independent experiments. p-values were assessed using a Student's t test

    Article Snippet: The following siRNAs were used: si-control A from Santa Cruz (sc-37007), si-Bcl-2 from Santa Cruz (sc-29214), si-Bcl-xL from Dharmacon (L-003458-00), si-Mcl-1 from Ambion (120644), si-Bim from Cell Signaling (6461), si-Puma from Dharmacon (L-004380-00), si-Myc from Santa Cruz (sc-29226), si-Foxo3A from Invitrogen (FOXO3a Validated Stealth DuoPak)

    Techniques: Transfection, Western Blot, Staining, FACS, Expressing

    Apoptotic signaling of evofosfamide as a single agent against MDA ‐ MB ‐231‐ TXSA cells. (A) MDA ‐ MB ‐231‐ TXSA ‐ TGL cells were seeded at 2 × 10 6 per T25 flask and were treated with evofosfamide at 50 μmol/L under normoxic (21% O 2 ) and hypoxic (1% O 2 ) conditions. Cells were then lysed and protein lysates were collected at 0, 6, 12, 24, and 48 h after treatment. Cell lysates were analyzed by polyacrylamide gel electrophoresis and transferred to PVDF membranes for immunodetection as described in Materials and Methods section. Evofosfamide treatment resulted in cleavage and prominent activation of the initiator caspase‐8 followed by caspase‐9, and caspase‐3 and the concomitant processing of BID and PARP protein, only at the later time point of 48 h, indicating that these changes were an effect rather than a cause of evofosfamide‐induced apoptosis. cIAP 1/2 levels were significantly reduced with evofosfamide treatment, under hypoxic conditions at the 24‐ and 48‐h time points, whereas the levels of XIAP , BAX , and Bcl‐2 remained unchanged. (B) A robust reduction in PI 3 kinase and phosphorylated AKT under hypoxic conditions after treatment with TH ‐302 at 24 and 48 h. However, no significant changes in the levels of either phosphorylated or total MAPK were detected following treatment.

    Journal: Cancer Medicine

    Article Title: Anticancer efficacy of the hypoxia‐activated prodrug evofosfamide (TH‐302) in osteolytic breast cancer murine models

    doi: 10.1002/cam4.599

    Figure Lengend Snippet: Apoptotic signaling of evofosfamide as a single agent against MDA ‐ MB ‐231‐ TXSA cells. (A) MDA ‐ MB ‐231‐ TXSA ‐ TGL cells were seeded at 2 × 10 6 per T25 flask and were treated with evofosfamide at 50 μmol/L under normoxic (21% O 2 ) and hypoxic (1% O 2 ) conditions. Cells were then lysed and protein lysates were collected at 0, 6, 12, 24, and 48 h after treatment. Cell lysates were analyzed by polyacrylamide gel electrophoresis and transferred to PVDF membranes for immunodetection as described in Materials and Methods section. Evofosfamide treatment resulted in cleavage and prominent activation of the initiator caspase‐8 followed by caspase‐9, and caspase‐3 and the concomitant processing of BID and PARP protein, only at the later time point of 48 h, indicating that these changes were an effect rather than a cause of evofosfamide‐induced apoptosis. cIAP 1/2 levels were significantly reduced with evofosfamide treatment, under hypoxic conditions at the 24‐ and 48‐h time points, whereas the levels of XIAP , BAX , and Bcl‐2 remained unchanged. (B) A robust reduction in PI 3 kinase and phosphorylated AKT under hypoxic conditions after treatment with TH ‐302 at 24 and 48 h. However, no significant changes in the levels of either phosphorylated or total MAPK were detected following treatment.

    Article Snippet: Immunodetection was performed overnight at 4°C in PBS/blocking reagent containing 0.1% Tween 20, using the following primary antibodies at the dilutions suggested by the manufacturer. mAb anticaspase‐8, mAb anticaspase‐3, pAb antibid, pAb anticaspase‐9, pAb PI3 kinase Class III, pAb phospho‐p44/42 MAPK, pAb p44/42 MAPK, pAb phospho‐Akt, and pAb Akt were purchased from Cell Signaling Technology (Beverly, MA), pAb anti‐inhibitor of apoptosis 1 (cIAP1), pAb anti‐inhibitor of apoptosis 2 (cIAP2), pAb anti‐XIAP from R & D systems, pAb anti‐BAX, pAb anti‐p53, pAb anti‐p21, and mAb anti‐Bcl‐2 were purchased from Santa Cruz Biotechnology (Dallas, Texas) and pAb anti‐poly‐(ADP‐Ribose) polymerase (PARP) from Roche Diagnostics.

    Techniques: Multiple Displacement Amplification, Polyacrylamide Gel Electrophoresis, Immunodetection, Activation Assay

    In the MCF-7 resistant cell variant (MCF-7 R ), Resv attenuates phosphorylation in S15 and S46 of p53 by dephosphorylation and deactivation of ATM. However, it activates kinases CK1, CHK2, and AMPK to induce phosphorylation of p53 in S20 (which is required to activate p53 in order to upregulate BAX and PUMA genes) and modifies the ratio between BCL-2/BAX expression. The BAX protein was increased while BCL-2 protein was decreased, restoring apoptosis and overcoming chemoresistance. On the other hand, the overexpression of BCL-2 in MCF-7 R cells after CDDP treatment maintains the chemoresistance and blocks apoptosis despite the phosphorylation of p53 in S15 and S46 and the upregulation of NOXA and PUMA .

    Journal: Nutrients

    Article Title: Induction of p53 Phosphorylation at Serine 20 by Resveratrol Is Required to Activate p53 Target Genes, Restoring Apoptosis in MCF-7 Cells Resistant to Cisplatin

    doi: 10.3390/nu10091148

    Figure Lengend Snippet: In the MCF-7 resistant cell variant (MCF-7 R ), Resv attenuates phosphorylation in S15 and S46 of p53 by dephosphorylation and deactivation of ATM. However, it activates kinases CK1, CHK2, and AMPK to induce phosphorylation of p53 in S20 (which is required to activate p53 in order to upregulate BAX and PUMA genes) and modifies the ratio between BCL-2/BAX expression. The BAX protein was increased while BCL-2 protein was decreased, restoring apoptosis and overcoming chemoresistance. On the other hand, the overexpression of BCL-2 in MCF-7 R cells after CDDP treatment maintains the chemoresistance and blocks apoptosis despite the phosphorylation of p53 in S15 and S46 and the upregulation of NOXA and PUMA .

    Article Snippet: The AMPK inhibitor Compound C (or dorsomorphin), the CK1 inhibitor D4476, the Chk2 inhibitor, anti-rabbit and anti-mouse secondary antibodies, mouse monoclonal anti-phospho-ATM (S1981), rabbit polyclonal anti-ATM, monoclonal anti-p53-HRP (DO-1), and monoclonal anti-BCL-2 were purchased from Santa Cruz Biotechnology (San Diego, CA, USA).

    Techniques: Variant Assay, De-Phosphorylation Assay, Expressing, Over Expression

    Resv promotes early dephosphorylation of ATM, inhibition of BCL-2, and upregulation of BAX. MCF-7 R cells were treated with CDDP (6 μM) with or without Resv (100 μM) for 6 h. ( A ) Total and phospho-ATM and ( B ) BCL-2 and BAX proteins were assessed by Western blot using antibodies directed against total ATM, a specific phosphorylated residue on S1981 of ATM, BCL-2, or BAX. MCF-7 cells with CDPP were used as positive control of ATM phosphorylation. ( C ) Densitometric analysis of BCL-2 and ( D ) BAX after β-actin normalization. ( E ) Ratio between BCL-2/BAX by t test. All results are presented as mean of three independent experiments ± SD. * p

    Journal: Nutrients

    Article Title: Induction of p53 Phosphorylation at Serine 20 by Resveratrol Is Required to Activate p53 Target Genes, Restoring Apoptosis in MCF-7 Cells Resistant to Cisplatin

    doi: 10.3390/nu10091148

    Figure Lengend Snippet: Resv promotes early dephosphorylation of ATM, inhibition of BCL-2, and upregulation of BAX. MCF-7 R cells were treated with CDDP (6 μM) with or without Resv (100 μM) for 6 h. ( A ) Total and phospho-ATM and ( B ) BCL-2 and BAX proteins were assessed by Western blot using antibodies directed against total ATM, a specific phosphorylated residue on S1981 of ATM, BCL-2, or BAX. MCF-7 cells with CDPP were used as positive control of ATM phosphorylation. ( C ) Densitometric analysis of BCL-2 and ( D ) BAX after β-actin normalization. ( E ) Ratio between BCL-2/BAX by t test. All results are presented as mean of three independent experiments ± SD. * p

    Article Snippet: The AMPK inhibitor Compound C (or dorsomorphin), the CK1 inhibitor D4476, the Chk2 inhibitor, anti-rabbit and anti-mouse secondary antibodies, mouse monoclonal anti-phospho-ATM (S1981), rabbit polyclonal anti-ATM, monoclonal anti-p53-HRP (DO-1), and monoclonal anti-BCL-2 were purchased from Santa Cruz Biotechnology (San Diego, CA, USA).

    Techniques: De-Phosphorylation Assay, Inhibition, Western Blot, Positive Control

    LC suppresses lung cancer cell growth via an apoptosis-independent mechanism. (a) Cells were treated with LC (4 μ g/mL) for 6 h, 12 h, and 24 h. Western blot assays were performed to determine the expression of Bcl-2, Bax, and p53 in H460 cells. GAPDH was used as a loading control. (b) Western blot assays were performed to determine the expression of cleaved caspase-3, cleaved caspase-9, and cytochrome C in H460 cells.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Capilliposide Isolated from Lysimachia capillipes Hemsl. Induces ROS Generation, Cell Cycle Arrest, and Apoptosis in Human Nonsmall Cell Lung Cancer Cell Lines

    doi: 10.1155/2014/497456

    Figure Lengend Snippet: LC suppresses lung cancer cell growth via an apoptosis-independent mechanism. (a) Cells were treated with LC (4 μ g/mL) for 6 h, 12 h, and 24 h. Western blot assays were performed to determine the expression of Bcl-2, Bax, and p53 in H460 cells. GAPDH was used as a loading control. (b) Western blot assays were performed to determine the expression of cleaved caspase-3, cleaved caspase-9, and cytochrome C in H460 cells.

    Article Snippet: The monoclonal antibody against Bcl-2 (#sc-492) was obtained from Santa Cruz.

    Techniques: Western Blot, Expressing

    ER stress responses is involved in apoptosis of Mtb infected macrophages from Sp110 transgenic cattle (A) Sp110 induces ER stress marker Hspa5. Macrophages isolated from control cattle and Sp110 transgenic cattle were infected with BCG (MOI 5:1) for 24 h. The expression of Sp110 and Hspa5 was determined by immunoblots. Con, macrophages from control cattle. TG, macrophages from Sp110 transgenic cattle. (B) Total RNA was extracted from macrophages of (A). The expression of HSPA8 and DDIT3 was determined by qPCR. (C) Macrophages were isolated from control cattle and Sp110 transgenic cattle, and then challenged with BCG (MOI 5:1) for 24 h. The TG cattle (4-PBA) sample was pretreated with 4-PBA for 1 h before Mtb infection. 4-PBA remained for the rest of the infection. Cell apoptosis was quantified by Annexin-V staining followed by flow cytometric analysis. (D) Macrophages isolated from Sp110 transgenic cattle were pretreated with 4-PBA for 1 h, and then infected with BCG (MOI 5:1) for 24 h and 48 h. 4-PBA remained for the rest of the infection. Intracellular mycobacterial number was determined by CFU counting at the indicated timepoints. (E) and (F) Sp110 inhibits BCL2 expression in bovine macrophages. Samples were prepared as (A), the protein levels of NCL and BCL2 were detected by immunoblots (E) and the mRNA of BCL2 was determined by qPCR (F). (G) Model of Sp110-mediated macrophage apoptosis. Sp110 competitively binds to Hspa5, which translocates Hspa5 into the nucleus to activate ER stress and initiate the apoptotic pathway. Moreover, Sp110 promotes protein degradation of Ncl and Rps3a, downregulating the anti-apoptotic Bcl2 expression and upregulating PARP activity, thus promoting apoptosis. Data are present as means ±SD of three independent experiments,* p

    Journal: Oncotarget

    Article Title: Sp110 enhances macrophage resistance to Mycobacterium tuberculosis via inducing endoplasmic reticulum stress and inhibiting anti-apoptotic factors

    doi: 10.18632/oncotarget.19300

    Figure Lengend Snippet: ER stress responses is involved in apoptosis of Mtb infected macrophages from Sp110 transgenic cattle (A) Sp110 induces ER stress marker Hspa5. Macrophages isolated from control cattle and Sp110 transgenic cattle were infected with BCG (MOI 5:1) for 24 h. The expression of Sp110 and Hspa5 was determined by immunoblots. Con, macrophages from control cattle. TG, macrophages from Sp110 transgenic cattle. (B) Total RNA was extracted from macrophages of (A). The expression of HSPA8 and DDIT3 was determined by qPCR. (C) Macrophages were isolated from control cattle and Sp110 transgenic cattle, and then challenged with BCG (MOI 5:1) for 24 h. The TG cattle (4-PBA) sample was pretreated with 4-PBA for 1 h before Mtb infection. 4-PBA remained for the rest of the infection. Cell apoptosis was quantified by Annexin-V staining followed by flow cytometric analysis. (D) Macrophages isolated from Sp110 transgenic cattle were pretreated with 4-PBA for 1 h, and then infected with BCG (MOI 5:1) for 24 h and 48 h. 4-PBA remained for the rest of the infection. Intracellular mycobacterial number was determined by CFU counting at the indicated timepoints. (E) and (F) Sp110 inhibits BCL2 expression in bovine macrophages. Samples were prepared as (A), the protein levels of NCL and BCL2 were detected by immunoblots (E) and the mRNA of BCL2 was determined by qPCR (F). (G) Model of Sp110-mediated macrophage apoptosis. Sp110 competitively binds to Hspa5, which translocates Hspa5 into the nucleus to activate ER stress and initiate the apoptotic pathway. Moreover, Sp110 promotes protein degradation of Ncl and Rps3a, downregulating the anti-apoptotic Bcl2 expression and upregulating PARP activity, thus promoting apoptosis. Data are present as means ±SD of three independent experiments,* p

    Article Snippet: Primary antibodies used were Hspa8 (1966-1), Pcna (2714-1), Rps3a (7944-1), Ybx1 (2397-1), Prdx1 (3688-1), Bag2 (2622-1) purchased from Epitomics (Burlingame, CA), Wwox (4045S), Eif4g2 (5169P), Mcm3 (4012S), Myc (2276S), Casp3 (9662S), Eif2α (9722S), P-Eif2α (9721S), Vav1 (2502S), CHOP (5554S) and Anxa2 (8235S) purchased from Cell Signaling Technology (Danvers, MA), Oasl1 (sc-98385), Hspa5 (sc-13968), Sp110 (sc-98365), Ncl (sc-13057), and Bcl2 (sc-492) purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX), FLAG (F1804-200ug, Sigma, Saint Louis, MO), Vcp (Ab109240, Abcam Cambridge, MA), Atp2a2 (AB54032-25ul, Sangon, Shanghai, China), and Actin (TransGen Biotech Co., Ltd, Beijing, China).

    Techniques: Infection, Transgenic Assay, Marker, Isolation, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Staining, Flow Cytometry, Activity Assay

    Sp110 promotes Ncl and Rps3a degradation (A) Overexpression of Sp110 in RAW264.7 cells decreased the protein levels of Ncl and Bcl2. The protein levels of Sp110, Ncl and Bcl2 in Uninfected and H37Ra infected (MOI 5:1, 24 h) RAW-Control or RAW-Sp110 cells were determined by immunoblotting. (B) and (C) RAW-Control and RAW-Sp110 cells were infected with H37Ra (MOI 5:1, 24 h), and uninfected cells were used as controls. Expression of Bcl2 and Ncl was monitored by qPCR. (D) Sp110 promotes Ncl protein degradation. RAW-Control and RAW-Sp110 cells were treated with protein synthesis Cycloheximide (10 μg/ml) or proteasome inhibitor MG132 (10 μM) for different periods of time. Expression changes of Ncl were determined by immunoblotting. The intensity of Ncl bands was quantified with ImageJ and normalized to the intensity of Actin. (E) Sp110 inhibits Bcl2 expression via downregulating Ncl. Expression of Ncl and Bcl2 in RAW-Control, empty vector transfected RAW-Sp110, and pCMV-Myc- Ncl transfected RAW-Sp110 cells was monitored by immunoblotting. (F) and (G) Knockdown of Ncl leads to the downregulation of both mRNA and protein levels of the Bcl2. RAW264.7 cells were transfected with siRNAs targeting Ncl or negative control siRNA for 48 h, and then the mRNA and protein expression of Ncl and Bcl2 was determined by qPCR and immunoblotting respectively. (H) Sp110 decreases the stability of the reporter gene that contains the ARE sequence of Bcl2 mRNA. RAW264.7 cells were cotransfected with psiCHECK2- Bcl2 ARE and pCDNA3.1-FLAG- Sp110 or pCMV-Myc- Ncl for 48 h followed by luciferase assays. (I) and (J) Expression of Rps3a in RAW-Control and RAW-Sp110 cells was determined by immunoblotting and qPCR respectively. (K) Sp110 promotes Rps3a protein degradation. RAW-Control and RAW-Sp110 cells were treated with protein synthesis Cycloheximide (10 μg/ml) for different periods of time. Expression changes of Rps3a were determined by immunoblotting. (L) Overexpression of Sp110 enhances PARP activity. Data are present as means ±SD of three independent experiments, **p

    Journal: Oncotarget

    Article Title: Sp110 enhances macrophage resistance to Mycobacterium tuberculosis via inducing endoplasmic reticulum stress and inhibiting anti-apoptotic factors

    doi: 10.18632/oncotarget.19300

    Figure Lengend Snippet: Sp110 promotes Ncl and Rps3a degradation (A) Overexpression of Sp110 in RAW264.7 cells decreased the protein levels of Ncl and Bcl2. The protein levels of Sp110, Ncl and Bcl2 in Uninfected and H37Ra infected (MOI 5:1, 24 h) RAW-Control or RAW-Sp110 cells were determined by immunoblotting. (B) and (C) RAW-Control and RAW-Sp110 cells were infected with H37Ra (MOI 5:1, 24 h), and uninfected cells were used as controls. Expression of Bcl2 and Ncl was monitored by qPCR. (D) Sp110 promotes Ncl protein degradation. RAW-Control and RAW-Sp110 cells were treated with protein synthesis Cycloheximide (10 μg/ml) or proteasome inhibitor MG132 (10 μM) for different periods of time. Expression changes of Ncl were determined by immunoblotting. The intensity of Ncl bands was quantified with ImageJ and normalized to the intensity of Actin. (E) Sp110 inhibits Bcl2 expression via downregulating Ncl. Expression of Ncl and Bcl2 in RAW-Control, empty vector transfected RAW-Sp110, and pCMV-Myc- Ncl transfected RAW-Sp110 cells was monitored by immunoblotting. (F) and (G) Knockdown of Ncl leads to the downregulation of both mRNA and protein levels of the Bcl2. RAW264.7 cells were transfected with siRNAs targeting Ncl or negative control siRNA for 48 h, and then the mRNA and protein expression of Ncl and Bcl2 was determined by qPCR and immunoblotting respectively. (H) Sp110 decreases the stability of the reporter gene that contains the ARE sequence of Bcl2 mRNA. RAW264.7 cells were cotransfected with psiCHECK2- Bcl2 ARE and pCDNA3.1-FLAG- Sp110 or pCMV-Myc- Ncl for 48 h followed by luciferase assays. (I) and (J) Expression of Rps3a in RAW-Control and RAW-Sp110 cells was determined by immunoblotting and qPCR respectively. (K) Sp110 promotes Rps3a protein degradation. RAW-Control and RAW-Sp110 cells were treated with protein synthesis Cycloheximide (10 μg/ml) for different periods of time. Expression changes of Rps3a were determined by immunoblotting. (L) Overexpression of Sp110 enhances PARP activity. Data are present as means ±SD of three independent experiments, **p

    Article Snippet: Primary antibodies used were Hspa8 (1966-1), Pcna (2714-1), Rps3a (7944-1), Ybx1 (2397-1), Prdx1 (3688-1), Bag2 (2622-1) purchased from Epitomics (Burlingame, CA), Wwox (4045S), Eif4g2 (5169P), Mcm3 (4012S), Myc (2276S), Casp3 (9662S), Eif2α (9722S), P-Eif2α (9721S), Vav1 (2502S), CHOP (5554S) and Anxa2 (8235S) purchased from Cell Signaling Technology (Danvers, MA), Oasl1 (sc-98385), Hspa5 (sc-13968), Sp110 (sc-98365), Ncl (sc-13057), and Bcl2 (sc-492) purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX), FLAG (F1804-200ug, Sigma, Saint Louis, MO), Vcp (Ab109240, Abcam Cambridge, MA), Atp2a2 (AB54032-25ul, Sangon, Shanghai, China), and Actin (TransGen Biotech Co., Ltd, Beijing, China).

    Techniques: Over Expression, Infection, Expressing, Real-time Polymerase Chain Reaction, Plasmid Preparation, Transfection, Negative Control, Sequencing, Luciferase, Activity Assay

    Metformin increased cell apoptosis as measured by p27, Bax, Bcl-2 and cleaved caspase-3 in MCF-7 cells. ( A ) Western blot of p27 and β-actin from MCF-7 cells that were treated with metformin (10 mM) for 24, 48 and 72 hours. ( B ) Western blot ratio analysis of p27 and β-actin. ( C ) Western blot of Bax, Bcl-2 and β-actin from MCF-7 cells that were treated with metformin (10 mM) for 24, 48 and 72 hours. ( D ) Western blot ratio analysis of Bax and β-actin, ( E ) Bcl-2 and β-actin, and ( F ) Bax and Bcl-2 at 24, 48 and 72 hours. ( G ) Western blot of cleaved caspase-3 and β-actin from MCF-7 cells that were treated with 10 mM metformin for 24, 48, and 72 hours. ( H ) Western blot ratio analysis of cleaved caspase-3 and β-actin at 24, 48, and 72 hours. * p

    Journal: PLoS ONE

    Article Title: Metformin Induces Apoptosis and Cell Cycle Arrest Mediated by Oxidative Stress, AMPK and FOXO3a in MCF-7 Breast Cancer Cells

    doi: 10.1371/journal.pone.0098207

    Figure Lengend Snippet: Metformin increased cell apoptosis as measured by p27, Bax, Bcl-2 and cleaved caspase-3 in MCF-7 cells. ( A ) Western blot of p27 and β-actin from MCF-7 cells that were treated with metformin (10 mM) for 24, 48 and 72 hours. ( B ) Western blot ratio analysis of p27 and β-actin. ( C ) Western blot of Bax, Bcl-2 and β-actin from MCF-7 cells that were treated with metformin (10 mM) for 24, 48 and 72 hours. ( D ) Western blot ratio analysis of Bax and β-actin, ( E ) Bcl-2 and β-actin, and ( F ) Bax and Bcl-2 at 24, 48 and 72 hours. ( G ) Western blot of cleaved caspase-3 and β-actin from MCF-7 cells that were treated with 10 mM metformin for 24, 48, and 72 hours. ( H ) Western blot ratio analysis of cleaved caspase-3 and β-actin at 24, 48, and 72 hours. * p

    Article Snippet: All primary antibodies were purchased from Cell Signaling Technologies (Danvers, MA, USA), except for Bax and Bcl-2 antibodies which were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Western Blot

    The anticancer effects of AgNPs on the PC-3 cells, a cytotoxic effects and b effects of the level of phosphorylated stat 3, bcl-2, survivin, and caspase-3

    Journal: Nanoscale Research Letters

    Article Title: Biosynthesis, Antibacterial Activity and Anticancer Effects Against Prostate Cancer (PC-3) Cells of Silver Nanoparticles Using Dimocarpus Longan Lour. Peel Extract

    doi: 10.1186/s11671-016-1511-9

    Figure Lengend Snippet: The anticancer effects of AgNPs on the PC-3 cells, a cytotoxic effects and b effects of the level of phosphorylated stat 3, bcl-2, survivin, and caspase-3

    Article Snippet: Bcl-2 antibody and all secondary antibodies were provided by Santa Cruz Biotechnology (Dallas, TX, USA).

    Techniques:

    The Bax:Bcl-2 ratio measured by Western blot analysis. Four treatment groups: control (CTL); 40 μM dexamethasone (DXM) for 8 h; 100 μM temozolomide (TMZ) for 6 h; pretreatment with 40 μM DXM for 2 h followed by 100 μM TMZ for 6 h. (A) A representative gel picture showing level of expression of Bax. (B) A representative gel picture showing level of expression of Bcl-2. (C) A representative gel picture showing level of expression of β-actin. (D) Densitometric analysis showing the Bax:Bcl-2 ratio in all treatment groups. Significant difference between CTL and TMZ treated cells was indicated by * ( P ≤ 0.05) and significant difference between TMZ treated cells and DXM plus TMZ treated cells was indicated by # ( P ≤ 0.05).

    Journal: Molecular Cancer

    Article Title: Dexamethasone protected human glioblastoma U87MG cells from temozolomide induced apoptosis by maintaining Bax:Bcl-2 ratio and preventing proteolytic activities

    doi: 10.1186/1476-4598-3-36

    Figure Lengend Snippet: The Bax:Bcl-2 ratio measured by Western blot analysis. Four treatment groups: control (CTL); 40 μM dexamethasone (DXM) for 8 h; 100 μM temozolomide (TMZ) for 6 h; pretreatment with 40 μM DXM for 2 h followed by 100 μM TMZ for 6 h. (A) A representative gel picture showing level of expression of Bax. (B) A representative gel picture showing level of expression of Bcl-2. (C) A representative gel picture showing level of expression of β-actin. (D) Densitometric analysis showing the Bax:Bcl-2 ratio in all treatment groups. Significant difference between CTL and TMZ treated cells was indicated by * ( P ≤ 0.05) and significant difference between TMZ treated cells and DXM plus TMZ treated cells was indicated by # ( P ≤ 0.05).

    Article Snippet: Bax and Bcl-2 monoclonal antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) were used to assess apoptotic threshold by determining the Bax:Bcl-2 ratio.

    Techniques: Western Blot, CTL Assay, Expressing

    GSK3β was downregulated by reduction of Bmi1 expression. (A) GBM cell lines U373 and Hs683 transduced with shRNA against Bmi1 (sh1061) or with scrambled shRNA (scr). Bmi1 downregulation induced downregulation of the following proteins: p-AKT, Nestin (in U373), Bcl2, and GSK3β. No effects are evident on the p16/p14 ARF , or p-ERK protein levels. (B) Scrambled control or U373 Bmi1-downregulated cells at subconfluency (sub-con) or confluency (con). Bmi1-downregulated cells show differentiated morphology (arrow: long, branched processes). (C) The glial fibrillary acidic protein (GFAP) levels in U373scr and U373sh1061 cells measured by flow cytometry. (D) Relative CNPase protein levels in U373scr and U373sh1061 measured by western blotting, quantified using ImageJ software and normalized to β-actin levels. (E) GSK3β mRNA values from a series of primary brain tumors (GBM and astrocytoma) and normal brain. (F) GSK3β protein is expressed in primary GBM. * P

    Journal: PLoS ONE

    Article Title: GSK3? Regulates Differentiation and Growth Arrest in Glioblastoma

    doi: 10.1371/journal.pone.0007443

    Figure Lengend Snippet: GSK3β was downregulated by reduction of Bmi1 expression. (A) GBM cell lines U373 and Hs683 transduced with shRNA against Bmi1 (sh1061) or with scrambled shRNA (scr). Bmi1 downregulation induced downregulation of the following proteins: p-AKT, Nestin (in U373), Bcl2, and GSK3β. No effects are evident on the p16/p14 ARF , or p-ERK protein levels. (B) Scrambled control or U373 Bmi1-downregulated cells at subconfluency (sub-con) or confluency (con). Bmi1-downregulated cells show differentiated morphology (arrow: long, branched processes). (C) The glial fibrillary acidic protein (GFAP) levels in U373scr and U373sh1061 cells measured by flow cytometry. (D) Relative CNPase protein levels in U373scr and U373sh1061 measured by western blotting, quantified using ImageJ software and normalized to β-actin levels. (E) GSK3β mRNA values from a series of primary brain tumors (GBM and astrocytoma) and normal brain. (F) GSK3β protein is expressed in primary GBM. * P

    Article Snippet: The following primary antibodies were used: anti-Bmi1 (Upstate), anti-β-catenin and anti-Nestin (Santa Cruz Biotechnology); anti-β-tubulin III and anti-GFAP (Sigma); anti-CNPase (Chemicon); anti-GSK3β (Cell Signaling); anti-Notch2 (Developmental Studies Hybridoma Bank); anti-Sox2 (R & D systems); anti-CD133 (Miltenyi Biotec); anti-Akt and phospho-Akt (Ser-473) (Millipore, Billerica MA, USA), anti-p16/p14, anti-Bcl2, anti-Erk and anti-phospho-Erk (Santa Cruz Biotechnology, Santa Cruz CA, USA), anti-Actin (Sigma-Aldrich, St. Louis, USA).

    Techniques: Expressing, Transduction, shRNA, Flow Cytometry, Cytometry, Western Blot, Software

    Figure 6. Adaptation to low Ca 2+ conditions reduces ER Ca 2+ levels similarly to Bcl-2 expression, and mimics the Bcl-2 dependent increase in NE permeability. (A) Typical traces of Fluo-4 fluorescence in HeLa cells stably expressing Bcl-2 (light

    Journal: Nucleus

    Article Title: Regulation of nuclear envelope permeability in cell death and survival

    doi: 10.4161/nucl.21982

    Figure Lengend Snippet: Figure 6. Adaptation to low Ca 2+ conditions reduces ER Ca 2+ levels similarly to Bcl-2 expression, and mimics the Bcl-2 dependent increase in NE permeability. (A) Typical traces of Fluo-4 fluorescence in HeLa cells stably expressing Bcl-2 (light

    Article Snippet: Immunocytochemical detection of m bcl-2 was performed using antibodies specific for murine bcl-2 (mouse monoclonal anti-bcl-2 clone 10C4, Santa Cruz), at a dilution of 1:500 in 10% NGS/PBS.

    Techniques: Expressing, Permeability, Fluorescence, Stable Transfection

    Figure 5. Bcl-2 localization at the nuclear membrane is sufficient to increase the size exclusion limit of the NE. (A) Confocal images of HeLa cells transiently expressing the permeability marker 4xCherry alone (control) and in combination with

    Journal: Nucleus

    Article Title: Regulation of nuclear envelope permeability in cell death and survival

    doi: 10.4161/nucl.21982

    Figure Lengend Snippet: Figure 5. Bcl-2 localization at the nuclear membrane is sufficient to increase the size exclusion limit of the NE. (A) Confocal images of HeLa cells transiently expressing the permeability marker 4xCherry alone (control) and in combination with

    Article Snippet: Immunocytochemical detection of m bcl-2 was performed using antibodies specific for murine bcl-2 (mouse monoclonal anti-bcl-2 clone 10C4, Santa Cruz), at a dilution of 1:500 in 10% NGS/PBS.

    Techniques: Expressing, Permeability, Marker

    Figure 4. Bcl-2 overexpression leads to increased nuclear envelope permeability. (A) Nuclear entry of 70 kD fluorescent dextran was assessed in HeLa and SW480 cells stably expressing murine and human bcl-2, respectively, using the Nuclear Permeability

    Journal: Nucleus

    Article Title: Regulation of nuclear envelope permeability in cell death and survival

    doi: 10.4161/nucl.21982

    Figure Lengend Snippet: Figure 4. Bcl-2 overexpression leads to increased nuclear envelope permeability. (A) Nuclear entry of 70 kD fluorescent dextran was assessed in HeLa and SW480 cells stably expressing murine and human bcl-2, respectively, using the Nuclear Permeability

    Article Snippet: Immunocytochemical detection of m bcl-2 was performed using antibodies specific for murine bcl-2 (mouse monoclonal anti-bcl-2 clone 10C4, Santa Cruz), at a dilution of 1:500 in 10% NGS/PBS.

    Techniques: Over Expression, Permeability, Stable Transfection, Expressing

    Evaluation of apoptosis in the zone of stasis after burns with HS management. Classic TUNEL staining indicated symbolic brown nuclei that were mainly located in the dermis of the interspace skin postburn (400× magnification)(A). Counting the apoptotic cells indicated a significant elevation in the burn controls over time, with a peak occurring at 72 h post burn. Except for the 6 h group, HS administration effectively decreased the number of apoptotic cells in the treatment groups at 24 h and 48 h (400× magnification)(A). Bcl-2 expression was downregulated after burn insult, while the protein expression of Bax and caspase-3 exhibited increasing trends over time (B-E). Nevertheless, HS seemed to reverse these of expression tendencies at various time points (B-E). The sample size was n = 5 for each group. The results were expressed as the mean±SEM. *p

    Journal: PLoS ONE

    Article Title: Beneficial Effects of Hydrogen-Rich Saline on Early Burn-Wound Progression in Rats

    doi: 10.1371/journal.pone.0124897

    Figure Lengend Snippet: Evaluation of apoptosis in the zone of stasis after burns with HS management. Classic TUNEL staining indicated symbolic brown nuclei that were mainly located in the dermis of the interspace skin postburn (400× magnification)(A). Counting the apoptotic cells indicated a significant elevation in the burn controls over time, with a peak occurring at 72 h post burn. Except for the 6 h group, HS administration effectively decreased the number of apoptotic cells in the treatment groups at 24 h and 48 h (400× magnification)(A). Bcl-2 expression was downregulated after burn insult, while the protein expression of Bax and caspase-3 exhibited increasing trends over time (B-E). Nevertheless, HS seemed to reverse these of expression tendencies at various time points (B-E). The sample size was n = 5 for each group. The results were expressed as the mean±SEM. *p

    Article Snippet: Subsequently, membranes were incubated in blocking buffer for two hours and incubated overnight at 4°C with the following primary antibodies: anti-TNF-α antibody (SC-1351, Santa Cruz Biotechnology, CA, USA), anti-IL-1β antibody (SC-23460, Santa Cruz Biotechnology, CA, USA), anti-IL-6 antibody (SC-1265, Santa Cruz Biotechnology, CA, USA), anti-IL-10 antibody (ab9969, Abcam, Cambridge, UK), anti-cleaved caspase-3 antibody (1:500)(SC-22171-R, Santa Cruz Biotechnology, CA, USA), anti-Bcl-2 antibody (1:800)(SC-783, Santa Cruz Biotechnology, CA, USA), anti-Bax antibody (1:500)(SC-493, Santa Cruz Biotechnology, CA, USA), anti-NF-κB p65 antibody (1:500)(ab16502, Abcam, Cambridge, UK), anti-p-Akt antibody (1:300)(SC-33437, Santa Cruz Biotechnology, CA, USA). β-actin (1:2000)(SC-47778, Santa Cruz Biotechnology, Santa Cruz, CA) was used as a control on the same membranes.

    Techniques: TUNEL Assay, Staining, Expressing

    Effect of co-treatment of breast cancer cell lines with 4-hydroxytamoxifen and erucin on PARP cleavage, levels of Bcl-2, Bax, survivin and ADRP. T47D ( a ) and MCF-7 ( b ) cells were treated with 5 μM erucin (ERN) and/or 0.5 μM 4-hydroxytamoxifen (4-OH-T). BT-474 ( c ) cells were treated with 5 μM erucin (ERN) and/or 1 μM 4-hydroxytamoxifen (4-OH-T). Blots were stripped and reprobed with anti-β-actin antibody to ensure equal protein loading. Results are plotted as mean ± SE from 3 independent experiments, *significantly different compared with single agent-treated samples or **significantly different compared with one of the single agent-treated samples by one-way ANOVA followed by Bonferroni’s multiple comparison test. Data for PARP refer to the faster migrating band marked as * and are given relative to samples treated with ERN alone. Blots shown are representative of at least three independent experiments

    Journal: European Journal of Nutrition

    Article Title: Sensitization of estrogen receptor-positive breast cancer cell lines to 4-hydroxytamoxifen by isothiocyanates present in cruciferous plants

    doi: 10.1007/s00394-015-0930-1

    Figure Lengend Snippet: Effect of co-treatment of breast cancer cell lines with 4-hydroxytamoxifen and erucin on PARP cleavage, levels of Bcl-2, Bax, survivin and ADRP. T47D ( a ) and MCF-7 ( b ) cells were treated with 5 μM erucin (ERN) and/or 0.5 μM 4-hydroxytamoxifen (4-OH-T). BT-474 ( c ) cells were treated with 5 μM erucin (ERN) and/or 1 μM 4-hydroxytamoxifen (4-OH-T). Blots were stripped and reprobed with anti-β-actin antibody to ensure equal protein loading. Results are plotted as mean ± SE from 3 independent experiments, *significantly different compared with single agent-treated samples or **significantly different compared with one of the single agent-treated samples by one-way ANOVA followed by Bonferroni’s multiple comparison test. Data for PARP refer to the faster migrating band marked as * and are given relative to samples treated with ERN alone. Blots shown are representative of at least three independent experiments

    Article Snippet: Antibodies against Bcl-2, Bax, survivin and ADRP were from Santa Cruz Biotechnology (Santa Cruz, CA), antibody against PARP was from Cell Signaling Technology (Danvers, MA) and anti-LC3 antibody was purchased from Medical and Biological Laboratories Co., Ltd. (Woburn, MA).

    Techniques:

    Effect of co-treatment of breast cancer cell lines with 4-hydroxytamoxifen and sulforaphane on PARP cleavage and levels of Bcl-2, Bax, survivin and ADRP. T47D ( a ) and MCF-7 ( b ) cells were treated with 5 μM sulforaphane (SFN), and/or 0.5 μM 4-hydroxytamoxifen (4-OH-T). BT-474 ( c ) cells were treated with 5 μM sulforaphane (SFN) and/or 1 μM 4-hydroxytamoxifen (4-OH-T). Blots were stripped and reprobed with anti-β-actin antibody to ensure equal protein loading. Results are plotted as mean ± SE from three independent experiments, *significantly different compared with single agent-treated samples or **significantly different compared with one of the single agent-treated samples by one-way ANOVA followed by Bonferroni’s multiple comparison test. Data for PARP refer to the faster migrating band marked as * and are given relative to samples treated with SFN alone. Blots shown are representative of at least three independent experiments

    Journal: European Journal of Nutrition

    Article Title: Sensitization of estrogen receptor-positive breast cancer cell lines to 4-hydroxytamoxifen by isothiocyanates present in cruciferous plants

    doi: 10.1007/s00394-015-0930-1

    Figure Lengend Snippet: Effect of co-treatment of breast cancer cell lines with 4-hydroxytamoxifen and sulforaphane on PARP cleavage and levels of Bcl-2, Bax, survivin and ADRP. T47D ( a ) and MCF-7 ( b ) cells were treated with 5 μM sulforaphane (SFN), and/or 0.5 μM 4-hydroxytamoxifen (4-OH-T). BT-474 ( c ) cells were treated with 5 μM sulforaphane (SFN) and/or 1 μM 4-hydroxytamoxifen (4-OH-T). Blots were stripped and reprobed with anti-β-actin antibody to ensure equal protein loading. Results are plotted as mean ± SE from three independent experiments, *significantly different compared with single agent-treated samples or **significantly different compared with one of the single agent-treated samples by one-way ANOVA followed by Bonferroni’s multiple comparison test. Data for PARP refer to the faster migrating band marked as * and are given relative to samples treated with SFN alone. Blots shown are representative of at least three independent experiments

    Article Snippet: Antibodies against Bcl-2, Bax, survivin and ADRP were from Santa Cruz Biotechnology (Santa Cruz, CA), antibody against PARP was from Cell Signaling Technology (Danvers, MA) and anti-LC3 antibody was purchased from Medical and Biological Laboratories Co., Ltd. (Woburn, MA).

    Techniques:

    Bcl-2 and Bax expression in VAT-39 cells. ( A ) Western blot analysis of Bcl-2 and Bax in 5 μM cisplatin- and 2 μM SAHA-treated VAT-39 cells. Isolated proteins (10 μg) were subjected to SDS-PAGE. Rabbit anti-Bcl-2 and mouse anti-Bax antibodies were incubated with the same membrane. ( B ) Protein bands were analyzed with densitometry, and the result is shown as the Bax/Bcl-2 ratio. * P

    Journal: Acta Histochemica et Cytochemica

    Article Title: The HDAC Inhibitor, SAHA, Combined with Cisplatin Synergistically Induces Apoptosis in Alpha-fetoprotein-producing Hepatoid Adenocarcinoma Cells

    doi: 10.1267/ahc.18044

    Figure Lengend Snippet: Bcl-2 and Bax expression in VAT-39 cells. ( A ) Western blot analysis of Bcl-2 and Bax in 5 μM cisplatin- and 2 μM SAHA-treated VAT-39 cells. Isolated proteins (10 μg) were subjected to SDS-PAGE. Rabbit anti-Bcl-2 and mouse anti-Bax antibodies were incubated with the same membrane. ( B ) Protein bands were analyzed with densitometry, and the result is shown as the Bax/Bcl-2 ratio. * P

    Article Snippet: The membranes were blocked with 5% nonfat skim milk in Tris-buffered saline with 0.1% Tween 20 (TBST; 20 mM Tris buffer, pH 7.6, and 150 mM NaCl) for 1 hr at room temperature and then incubated overnight with primary antibody against Bcl-2 (N-19, 1:500), Bax (B-9, 1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), H3K9ac, H3K14ac, H3K18ac, H3K27ac (1:1,000), cleaved caspase-3 (1:500, Cell Signaling Technology), or β-actin (AC-15, 1:400,000 diluted, Sigma-Aldrich).

    Techniques: Expressing, Western Blot, Isolation, SDS Page, Incubation

    Bcl‐2 in ischemic brains of rats detected by immunohistochemistry in the ( A ) hippocampus and ( B ) cortex. The presence of Bcl‐2 was indicated by a brown peroxidase reaction product at ×200. Error bars are the SEM. * P

    Journal: CNS Neuroscience & Therapeutics

    Article Title: A Neuroprotective Mechanism of YGY‐E in Cerebral Ischemic Injury in Rats

    doi: 10.1111/j.1755-5949.2011.00277.x

    Figure Lengend Snippet: Bcl‐2 in ischemic brains of rats detected by immunohistochemistry in the ( A ) hippocampus and ( B ) cortex. The presence of Bcl‐2 was indicated by a brown peroxidase reaction product at ×200. Error bars are the SEM. * P

    Article Snippet: Sections were incubated with the primary antibody anti‐Bcl‐2 (Santa Cruz Biotechnology, CA, USA) at 37°C for 1 h and the secondary antibody biotinylated goat anti‐rat IgG (Santa Cruz) for 20 min. Bcl‐2 positive cells were identified by a brown peroxidase reaction product after incubation with an avidin–horseradish enzyme complex for 20 min at 37°C using an ABC Kit (Sino‐American Biotechnology Company, MA, USA) with DAB (Sino‐American Biotechnology Company, MA, USA) as the chromogen.

    Techniques: Immunohistochemistry

    Bcl‐2 expression in ischemic brains of rats by Western blot analysis in the ( A ) hippocampus and ( B ) cortex. Relative Bcl‐2 (%) was calculated as the density of the Bcl‐2 gel band/the density of the GAPDH gel band. Error bars are the SEM. * P

    Journal: CNS Neuroscience & Therapeutics

    Article Title: A Neuroprotective Mechanism of YGY‐E in Cerebral Ischemic Injury in Rats

    doi: 10.1111/j.1755-5949.2011.00277.x

    Figure Lengend Snippet: Bcl‐2 expression in ischemic brains of rats by Western blot analysis in the ( A ) hippocampus and ( B ) cortex. Relative Bcl‐2 (%) was calculated as the density of the Bcl‐2 gel band/the density of the GAPDH gel band. Error bars are the SEM. * P

    Article Snippet: Sections were incubated with the primary antibody anti‐Bcl‐2 (Santa Cruz Biotechnology, CA, USA) at 37°C for 1 h and the secondary antibody biotinylated goat anti‐rat IgG (Santa Cruz) for 20 min. Bcl‐2 positive cells were identified by a brown peroxidase reaction product after incubation with an avidin–horseradish enzyme complex for 20 min at 37°C using an ABC Kit (Sino‐American Biotechnology Company, MA, USA) with DAB (Sino‐American Biotechnology Company, MA, USA) as the chromogen.

    Techniques: Expressing, Western Blot

    Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL), Bax, B-cell lymphoma 2-associated X protein (Bax), and B-cell lymphoma 2 (Bcl-2) staining in the penumbra, and the effects of cilostazol (CZL) after 96 hours in Long-Evans Tokushima Otsuka (LETO) and Otsuka Long-Evans Tokushima Fatty (OLETF) rats. The number of apoptotic TUNEL-positive cells (with darkly stained nuclei or nuclear fragments with a cytoplasmic halo) was reduced by CZL. Strong Bax protein expression was observed both in the penumbra and in areas distant from the acute ischemic lesion, and was suppressed by CZL. Immunolabeling for Bcl-2 near the infarct area was increased by CZL (bar=200 μ m, × 200 magnification). CMC, carboxymethyl cellulose.

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: Cilostazol minimizes venous ischemic injury in diabetic and normal rats

    doi: 10.1038/jcbfm.2011.47

    Figure Lengend Snippet: Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL), Bax, B-cell lymphoma 2-associated X protein (Bax), and B-cell lymphoma 2 (Bcl-2) staining in the penumbra, and the effects of cilostazol (CZL) after 96 hours in Long-Evans Tokushima Otsuka (LETO) and Otsuka Long-Evans Tokushima Fatty (OLETF) rats. The number of apoptotic TUNEL-positive cells (with darkly stained nuclei or nuclear fragments with a cytoplasmic halo) was reduced by CZL. Strong Bax protein expression was observed both in the penumbra and in areas distant from the acute ischemic lesion, and was suppressed by CZL. Immunolabeling for Bcl-2 near the infarct area was increased by CZL (bar=200 μ m, × 200 magnification). CMC, carboxymethyl cellulose.

    Article Snippet: Endogenous peroxidase activity was blocked by incubation in 0.3% hydrogen peroxide (H2 O2 ), and the sections were incubated overnight with mouse monoclonal antibodies directed against smooth muscle actin (SMA; Santa Cruz Biochemicals, Santa Cruz, CA, USA; 1:100 dilution), B-cell lymphoma 2-associated X protein (Bax; SC7480; Santa Cruz Biochemicals; 1:500 dilution), B-cell lymphoma 2 (Bcl-2; SC7480; Santa Cruz Biochemicals; 1:300 dilution), and advanced glycation end products (AGEs; 6D12; TransGenic Inc., Kumamoto, Japan; 1:20 dilution).

    Techniques: End Labeling, TUNEL Assay, Staining, Expressing, Immunolabeling

    Infarct volume and rat es of TUNEL-, Bax-, and Bcl-2-positive cells. The venous infarct volume was reduced by cilostazol in the Long-Evans Tokushima Otsuka (LETO) rats. In the Otsuka Long-Evans Tokushima Fatty (OLTEF) rats, the infarct volume increased up to 48 hours after two-vein occlusion (2VO), but was significantly reduced by cilostazol after 96 hours. The percentage of TUNEL-positive cells was higher, but was reduced significantly after 96 hours, in the OLETF rats, while it was significantly reduced by cilostazol in the LETO rats. The percentage of Bax-positive cells showed the same pattern. The percentage of Bcl-2-positive cells near the infarct area significantly increased 48 hours after cilostazol administration in both the LETO and the OLETF rats. The data for the cilostazol-administered groups are represented by dark bars and the data for the carboxymethyl cellulose (CMC)-administered groups are represented by light bars. TUNEL, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling; Bax, B-cell lymphoma 2-associated X protein; Bcl-2, B-cell lymphoma 2.

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: Cilostazol minimizes venous ischemic injury in diabetic and normal rats

    doi: 10.1038/jcbfm.2011.47

    Figure Lengend Snippet: Infarct volume and rat es of TUNEL-, Bax-, and Bcl-2-positive cells. The venous infarct volume was reduced by cilostazol in the Long-Evans Tokushima Otsuka (LETO) rats. In the Otsuka Long-Evans Tokushima Fatty (OLTEF) rats, the infarct volume increased up to 48 hours after two-vein occlusion (2VO), but was significantly reduced by cilostazol after 96 hours. The percentage of TUNEL-positive cells was higher, but was reduced significantly after 96 hours, in the OLETF rats, while it was significantly reduced by cilostazol in the LETO rats. The percentage of Bax-positive cells showed the same pattern. The percentage of Bcl-2-positive cells near the infarct area significantly increased 48 hours after cilostazol administration in both the LETO and the OLETF rats. The data for the cilostazol-administered groups are represented by dark bars and the data for the carboxymethyl cellulose (CMC)-administered groups are represented by light bars. TUNEL, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling; Bax, B-cell lymphoma 2-associated X protein; Bcl-2, B-cell lymphoma 2.

    Article Snippet: Endogenous peroxidase activity was blocked by incubation in 0.3% hydrogen peroxide (H2 O2 ), and the sections were incubated overnight with mouse monoclonal antibodies directed against smooth muscle actin (SMA; Santa Cruz Biochemicals, Santa Cruz, CA, USA; 1:100 dilution), B-cell lymphoma 2-associated X protein (Bax; SC7480; Santa Cruz Biochemicals; 1:500 dilution), B-cell lymphoma 2 (Bcl-2; SC7480; Santa Cruz Biochemicals; 1:300 dilution), and advanced glycation end products (AGEs; 6D12; TransGenic Inc., Kumamoto, Japan; 1:20 dilution).

    Techniques: TUNEL Assay, End Labeling

    Stable expression of human Bcl-2 significantly reduces NS1-induced cell death. (A) Representative Bcl-2-stable transfectants of both KLNS and ULNS cells were detected with a Bcl-2 MAb, Bcl-2 100, by immunoblotting with 5 × 10 6 cells in the presence or absence of IPTG. The position of the Bcl-2 protein corresponding to approximately 26 kDa is indicated. (B and C) The kinetics of Bcl-2 inhibition of NS1-induced cell death was measured by trypan blue exclusion for both KLNS (B) and ULNS (C) cells with 5 × 10 5 cells for each cell line. The experiments were performed twice, with no significant difference between results.

    Journal: Journal of Virology

    Article Title: Human Parvovirus B19 Nonstructural (NS1) Protein Induces Apoptosis in Erythroid Lineage Cells

    doi:

    Figure Lengend Snippet: Stable expression of human Bcl-2 significantly reduces NS1-induced cell death. (A) Representative Bcl-2-stable transfectants of both KLNS and ULNS cells were detected with a Bcl-2 MAb, Bcl-2 100, by immunoblotting with 5 × 10 6 cells in the presence or absence of IPTG. The position of the Bcl-2 protein corresponding to approximately 26 kDa is indicated. (B and C) The kinetics of Bcl-2 inhibition of NS1-induced cell death was measured by trypan blue exclusion for both KLNS (B) and ULNS (C) cells with 5 × 10 5 cells for each cell line. The experiments were performed twice, with no significant difference between results.

    Article Snippet: Human Bcl-2 was detected with Bcl-2(100) mouse MAb (Santa Cruz).

    Techniques: Expressing, Inhibition

    The schematic diagram illustrates the possible neuroprotective pathways in the brain under oral B401 treatment. Oral B401 treatment may improve neuroprotection in R6/2 mice via enhancing anti-oxidative stress (marked by SOD2) and anti-apoptosis (marked by Bcl-2), while suppressing mutant huntingtin aggregation, ROS production, ER stress-related apoptosis (marked by calpain), and mitochondrial dysfunction-related apoptosis (marked by Bax and caspase 3) in the brain. Abbreviations: Bax, Bcl-2-associated X protein; Bcl-2, B-cell lymphoma 2; ER, endoplasmic reticulum; mHtt, mutant huntingtin; ROS, reactive oxygen species; SOD2, superoxide dismutase 2.

    Journal: Clinical Interventions in Aging

    Article Title: Oral treatment with the herbal formula B401 protects against aging-dependent neurodegeneration by attenuating oxidative stress and apoptosis in the brain of R6/2 mice

    doi: 10.2147/CIA.S93819

    Figure Lengend Snippet: The schematic diagram illustrates the possible neuroprotective pathways in the brain under oral B401 treatment. Oral B401 treatment may improve neuroprotection in R6/2 mice via enhancing anti-oxidative stress (marked by SOD2) and anti-apoptosis (marked by Bcl-2), while suppressing mutant huntingtin aggregation, ROS production, ER stress-related apoptosis (marked by calpain), and mitochondrial dysfunction-related apoptosis (marked by Bax and caspase 3) in the brain. Abbreviations: Bax, Bcl-2-associated X protein; Bcl-2, B-cell lymphoma 2; ER, endoplasmic reticulum; mHtt, mutant huntingtin; ROS, reactive oxygen species; SOD2, superoxide dismutase 2.

    Article Snippet: By using the heat-induced epitope retrieval method, brain tissue sections were separately stained at room temperature for 1 hour with antibodies of mutant huntingtin (mHtt) (Abcam Inc., Cambridge, MA, USA), SOD2 (Cell Signaling Technology Inc., Danvers, MA, USA), B-cell lymphoma 2 (Bcl-2) (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), Bcl-2 associated X protein (Bax) (Thermo Fisher Scientific Inc., Waltham, MA, USA), calpain (Cell Signaling Technology Inc.), and caspase 3 (Cell Signaling Technology Inc.).

    Techniques: Mouse Assay, Mutagenesis

    Molecular events underlying apoptosis in MIAPaCa-2 cells treated with MGDG. a Cytochrome c in mitochondria, b Cytochrome c in the cytosol, c PARP and cleaved PARP, c Pro caspase-3 and cleaved caspase-3, e Bax and f Bcl-2 levels were determined by western blotting. Protein levels were evaluated relative to HSP-60 or actin. * P

    Journal: Radiation Oncology (London, England)

    Article Title: MGDG extracted from spinach enhances the cytotoxicity of radiation in pancreatic cancer cells

    doi: 10.1186/s13014-016-0729-0

    Figure Lengend Snippet: Molecular events underlying apoptosis in MIAPaCa-2 cells treated with MGDG. a Cytochrome c in mitochondria, b Cytochrome c in the cytosol, c PARP and cleaved PARP, c Pro caspase-3 and cleaved caspase-3, e Bax and f Bcl-2 levels were determined by western blotting. Protein levels were evaluated relative to HSP-60 or actin. * P

    Article Snippet: Proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then transferred to nitrocellulose membranes that were blocked with 5% non-fat milk in PBS and probed overnight at 4 °C with primary antibodies against the following proteins: actin (Santa Cruz Biotechnology, Dallas, TX, USA), poly (ADP-ribose) polymerase (PARP) (Cell Signaling Technology, Danvers, MA, USA), caspase-3 (Cell Signaling Technology), pro-caspase-3 (GeneTex, Irvine, CA, USA), B cell lymphoma (Bcl)-2 (Santa Cruz Biotechnology), and Bcl-2-associated X protein (Bax) (Santa Cruz Biotechnology).

    Techniques: Western Blot

    Increase in chemotherapeutic drug-induced apoptosis by miR-218. SGC7901 cells were transfected with miR-218 mimic or the negative control and incubated 48 h with (A) ADM or (B) L-OHP. Proportions of apoptotic cells under the different conditions were determined using flow cytometry. Expression levels of Bcl-2 (C) and Bax (D) in SGC7901 cells were examined 48 h after transfection. Results are shown from three independent experiments. * P

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: miR-218 inhibits multidrug resistance (MDR) of gastric cancer cells by targeting Hedgehog/smoothened

    doi:

    Figure Lengend Snippet: Increase in chemotherapeutic drug-induced apoptosis by miR-218. SGC7901 cells were transfected with miR-218 mimic or the negative control and incubated 48 h with (A) ADM or (B) L-OHP. Proportions of apoptotic cells under the different conditions were determined using flow cytometry. Expression levels of Bcl-2 (C) and Bax (D) in SGC7901 cells were examined 48 h after transfection. Results are shown from three independent experiments. * P

    Article Snippet: The membrane was probed with primary monoclonal antibodies against Bcl-2 (1:200; Santa Cruz Biotechnology, CA, USA), Bax (1:100; Santa Cruz), P-gp (1:200; Santa Cruz), and SMO (1:500, Cell Signaling Technology, Boston, USA).

    Techniques: Transfection, Negative Control, Incubation, Flow Cytometry, Cytometry, Expressing

    Effect of PD on the expression of apoptosis-related proteins. (A) A549 human lung cancer cells were treated with increasing concentrations of PD for 24 and 48 h. Expression levels of Bcl-2 and Bax were detected by western blot analysis and β-actin was used as a control. (B and C) Bcl-2 and (D and E) Bax protein bands were quantitated by densitometry, and the data are presented as the mean ± SD from three experiments. * P

    Journal: Oncology Letters

    Article Title: Polydatin inhibits growth of lung cancer cells by inducing apoptosis and causing cell cycle arrest

    doi: 10.3892/ol.2013.1696

    Figure Lengend Snippet: Effect of PD on the expression of apoptosis-related proteins. (A) A549 human lung cancer cells were treated with increasing concentrations of PD for 24 and 48 h. Expression levels of Bcl-2 and Bax were detected by western blot analysis and β-actin was used as a control. (B and C) Bcl-2 and (D and E) Bax protein bands were quantitated by densitometry, and the data are presented as the mean ± SD from three experiments. * P

    Article Snippet: Primary antibodies against Bcl-2, Bax and cyclin D1 and secondary antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

    Techniques: Expressing, Western Blot

    Western blot analysis. Serine phosphorylation and Bcl-2 expression in spleen cells cultured in the presence and absence of T-CHO. (a) Serine phosphorylation; (b) Bcl-2 expression after 24 hr in culture and 96 hr in culture. Lanes 1 and 5, spleen cells from pre-exposed mice, cultured in the presence of T-CHO; lanes 2 and 4, cells from T-CHO pre-exposed mice, cultured in the absence T-CHO; lanes 3 and 7, cells from naive mice cultured in the presence of T-CHO and lanes 4 and 8, cells from naive mice cultured in the absence of T-CHO.

    Journal: Immunology

    Article Title: Induction of immunoglobulin G1, interleukin-6 and interleukin-10 by Taenia crassiceps metacestode carbohydrates

    doi: 10.1046/j.1365-2567.2002.01519.x

    Figure Lengend Snippet: Western blot analysis. Serine phosphorylation and Bcl-2 expression in spleen cells cultured in the presence and absence of T-CHO. (a) Serine phosphorylation; (b) Bcl-2 expression after 24 hr in culture and 96 hr in culture. Lanes 1 and 5, spleen cells from pre-exposed mice, cultured in the presence of T-CHO; lanes 2 and 4, cells from T-CHO pre-exposed mice, cultured in the absence T-CHO; lanes 3 and 7, cells from naive mice cultured in the presence of T-CHO and lanes 4 and 8, cells from naive mice cultured in the absence of T-CHO.

    Article Snippet: For Bcl-2 determination, a polyclonal antibody reactive with human and mouse Bcl-2 (Santa Cruz Biotechnology, Santa Cruz, CA, sc-783) was used.

    Techniques: Western Blot, Expressing, Cell Culture, Mouse Assay

    Prediction and experimental validation of GBM cell line sensitization to TRAIL or TMZ treatment by ABT-737 A. A movement vector (red) was calculated by combining the weightings for the (Bcl-2+Bcl-xL+Mcl-1) functional group in the first 3 PCs. (i) and (ii) provide different perspectives on the vectors, compatible with 3D PC spaces for TMZ or TRAIL responsiveness. B. 3D PC spaces for TMZ and TRAIL responsiveness with movement vectors applied to test cell lines. The length of the movement vectors was calculated from the respective Bcl-2 and Bcl-xL content of the cell lines. Movement of treatment resistant cell lines towards regions populated by cell lines with higher treatment responsiveness predicts sensitization by the Bcl-2/Bcl-xL antagonist ABT-737. C, D, E. Experimental validation of predictions. Cell viability was measured for single and combination treatments. (C) Cells were treated with TMZ (150 μM) ± ABT-737 (1 μM) for 48 h. (D) Cells were treated with TRAIL (100 ng/ml) ± ABT-737 (1 μM) for 48 h. Data show cell viability (mean ± SEM from n = 3 independent experiments) in relation to untreated controls (100%). One-way ANOVA with Bonferroni comparisons was performed for statistical analysis. **p

    Journal: Oncotarget

    Article Title: Predicting the cell death responsiveness and sensitization of glioma cells to TRAIL and temozolomide

    doi: 10.18632/oncotarget.10973

    Figure Lengend Snippet: Prediction and experimental validation of GBM cell line sensitization to TRAIL or TMZ treatment by ABT-737 A. A movement vector (red) was calculated by combining the weightings for the (Bcl-2+Bcl-xL+Mcl-1) functional group in the first 3 PCs. (i) and (ii) provide different perspectives on the vectors, compatible with 3D PC spaces for TMZ or TRAIL responsiveness. B. 3D PC spaces for TMZ and TRAIL responsiveness with movement vectors applied to test cell lines. The length of the movement vectors was calculated from the respective Bcl-2 and Bcl-xL content of the cell lines. Movement of treatment resistant cell lines towards regions populated by cell lines with higher treatment responsiveness predicts sensitization by the Bcl-2/Bcl-xL antagonist ABT-737. C, D, E. Experimental validation of predictions. Cell viability was measured for single and combination treatments. (C) Cells were treated with TMZ (150 μM) ± ABT-737 (1 μM) for 48 h. (D) Cells were treated with TRAIL (100 ng/ml) ± ABT-737 (1 μM) for 48 h. Data show cell viability (mean ± SEM from n = 3 independent experiments) in relation to untreated controls (100%). One-way ANOVA with Bonferroni comparisons was performed for statistical analysis. **p

    Article Snippet: The following primary antibodies were used: rabbit polyclonal Apaf-1 (Cat# 16941, Millipore); rabbit polyclonal Bak (Cat# sc-832, Santa Cruz); rabbit polyclonal Bax (Cat# 06-499, Upstate Biotechnology); mouse monoclonal Bcl-2 (Cat# sc-509, Santa Cruz); mouse monoclonal Bcl-xL (Cat# sc-8392); rabbit polyclonal Bid (Cat# 2002, Cell Signaling); rabbit monoclonal Bim (Cat# 2933, Cell Signaling); rabbit polyclonal caspase-3 (Cat# 9662, Cell Signaling); mouse monoclonal caspase-8 (Cat# ALX-804-242, Enzo Life Sciences); rabbit polyclonal caspase-9 (Cat# 9502, Cell Signaling); mouse monoclonal cFLIP (Cat# ALX-804-428, Enzo Life Sciences); mouse monoclonal FADD (Cat# 610399, BD Biosciences); mouse monoclonal Mcl-1 (Cat# 559027, BD Biosciences); mouse monoclonal Noxa (Cat# 13654, Abcam); rabbit polyclonal PUMA (Cat# 3041, ProSci Incorporated); mouse monoclonal SMAC/DIABLO (Cat# 2954, Cell Signaling); and mouse monoclonal XIAP (Cat# 610763, BD Biosciences); and mouse monoclonal β-actin (Cat# A5441, Sigma-Aldrich).

    Techniques: Plasmid Preparation, Functional Assay

    Western blotting studies. ( A ). Western blotting analysis of pAKT, tAKT, PCNA, MMP9, Bcl-2, Bax, p53, cleaved caspase-3 with β-actin as an internal reference. ( B ). Quantification of the Western blotting results. Data was presented as the mean ± SD. *p

    Journal: Drug Design, Development and Therapy

    Article Title: Hongjingtian Injection Inhibits Proliferation and Migration and Promotes Apoptosis in High Glucose-Induced Vascular Smooth Muscle Cells

    doi: 10.2147/DDDT.S220719

    Figure Lengend Snippet: Western blotting studies. ( A ). Western blotting analysis of pAKT, tAKT, PCNA, MMP9, Bcl-2, Bax, p53, cleaved caspase-3 with β-actin as an internal reference. ( B ). Quantification of the Western blotting results. Data was presented as the mean ± SD. *p

    Article Snippet: Mouse anti-Bcl-2 (Sc-7382) was obtained from Santa Cruz Biotechnology(Dallas, USA); rabbit anti-MMP9(Ep1254) was obtained from Abcam(Cambridge, UK); rabbit anti-cleaved caspase-3 (Asp175), rabbit anti-AKT (C67E7), rabbit anti-phospho-AKT (Ser473), rabbit anti-PCNA (D3H8P) and mouse anti-p53(1c12) were obtained from Cell Signaling Technology (Danvers, MA, USA); and mouse anti-Bax (Gb11007) was obtained from Servicebio (Wuhan, China).

    Techniques: Western Blot

    Peptide 2 Reverses Bcl-2’s Inhibition of IP3R Channel Opening In Vitro

    Journal:

    Article Title: Targeting Bcl-2-IP3 Receptor Interaction to Reverse Bcl-2's Inhibition of Apoptotic Calcium Signals

    doi: 10.1016/j.molcel.2008.06.014

    Figure Lengend Snippet: Peptide 2 Reverses Bcl-2’s Inhibition of IP3R Channel Opening In Vitro

    Article Snippet: The following antibodies were used: anti-human Bcl-2 (BD Biosciences), anti-mouse Bcl-2 (Santa Cruz Biotechnology), anti-IP3R type 1 (Calbiochem), anti-Bim (Sigma), anti-Bax (BD PharMingen), anti-GST (Amersham Biosciences), and anti-actin (Sigma).

    Techniques: Inhibition, In Vitro

    Peptide 2 Enhances Anti-CD3-Induced Apoptosis in Bcl-2-Positive Cells

    Journal:

    Article Title: Targeting Bcl-2-IP3 Receptor Interaction to Reverse Bcl-2's Inhibition of Apoptotic Calcium Signals

    doi: 10.1016/j.molcel.2008.06.014

    Figure Lengend Snippet: Peptide 2 Enhances Anti-CD3-Induced Apoptosis in Bcl-2-Positive Cells

    Article Snippet: The following antibodies were used: anti-human Bcl-2 (BD Biosciences), anti-mouse Bcl-2 (Santa Cruz Biotechnology), anti-IP3R type 1 (Calbiochem), anti-Bim (Sigma), anti-Bax (BD PharMingen), anti-GST (Amersham Biosciences), and anti-actin (Sigma).

    Techniques:

    Endogenous Bcl-2 Inhibits Anti-CD3-Induced Ca 2+ Elevation in Jurkat Cells

    Journal:

    Article Title: Targeting Bcl-2-IP3 Receptor Interaction to Reverse Bcl-2's Inhibition of Apoptotic Calcium Signals

    doi: 10.1016/j.molcel.2008.06.014

    Figure Lengend Snippet: Endogenous Bcl-2 Inhibits Anti-CD3-Induced Ca 2+ Elevation in Jurkat Cells

    Article Snippet: The following antibodies were used: anti-human Bcl-2 (BD Biosciences), anti-mouse Bcl-2 (Santa Cruz Biotechnology), anti-IP3R type 1 (Calbiochem), anti-Bim (Sigma), anti-Bax (BD PharMingen), anti-GST (Amersham Biosciences), and anti-actin (Sigma).

    Techniques:

    Bcl-2-IP3R Interaction Mapping

    Journal:

    Article Title: Targeting Bcl-2-IP3 Receptor Interaction to Reverse Bcl-2's Inhibition of Apoptotic Calcium Signals

    doi: 10.1016/j.molcel.2008.06.014

    Figure Lengend Snippet: Bcl-2-IP3R Interaction Mapping

    Article Snippet: The following antibodies were used: anti-human Bcl-2 (BD Biosciences), anti-mouse Bcl-2 (Santa Cruz Biotechnology), anti-IP3R type 1 (Calbiochem), anti-Bim (Sigma), anti-Bax (BD PharMingen), anti-GST (Amersham Biosciences), and anti-actin (Sigma).

    Techniques:

    An IP3R Peptide Inhibits Bcl-2-IP3R Interaction but Not Bim-Bcl-2 Interaction

    Journal:

    Article Title: Targeting Bcl-2-IP3 Receptor Interaction to Reverse Bcl-2's Inhibition of Apoptotic Calcium Signals

    doi: 10.1016/j.molcel.2008.06.014

    Figure Lengend Snippet: An IP3R Peptide Inhibits Bcl-2-IP3R Interaction but Not Bim-Bcl-2 Interaction

    Article Snippet: The following antibodies were used: anti-human Bcl-2 (BD Biosciences), anti-mouse Bcl-2 (Santa Cruz Biotechnology), anti-IP3R type 1 (Calbiochem), anti-Bim (Sigma), anti-Bax (BD PharMingen), anti-GST (Amersham Biosciences), and anti-actin (Sigma).

    Techniques:

    Peptide 2 Reverses Bcl-2’s Inhibition of Anti-CD3-Induced Ca 2+ Elevation

    Journal:

    Article Title: Targeting Bcl-2-IP3 Receptor Interaction to Reverse Bcl-2's Inhibition of Apoptotic Calcium Signals

    doi: 10.1016/j.molcel.2008.06.014

    Figure Lengend Snippet: Peptide 2 Reverses Bcl-2’s Inhibition of Anti-CD3-Induced Ca 2+ Elevation

    Article Snippet: The following antibodies were used: anti-human Bcl-2 (BD Biosciences), anti-mouse Bcl-2 (Santa Cruz Biotechnology), anti-IP3R type 1 (Calbiochem), anti-Bim (Sigma), anti-Bax (BD PharMingen), anti-GST (Amersham Biosciences), and anti-actin (Sigma).

    Techniques: Inhibition

    FRET Detection of Bcl-2-IP3R Interaction

    Journal:

    Article Title: Targeting Bcl-2-IP3 Receptor Interaction to Reverse Bcl-2's Inhibition of Apoptotic Calcium Signals

    doi: 10.1016/j.molcel.2008.06.014

    Figure Lengend Snippet: FRET Detection of Bcl-2-IP3R Interaction

    Article Snippet: The following antibodies were used: anti-human Bcl-2 (BD Biosciences), anti-mouse Bcl-2 (Santa Cruz Biotechnology), anti-IP3R type 1 (Calbiochem), anti-Bim (Sigma), anti-Bax (BD PharMingen), anti-GST (Amersham Biosciences), and anti-actin (Sigma).

    Techniques:

    Effects of EpCAM mAb on the induction of apoptosis in colon cancer cells analyzed by DAPI and TUNEL staining. Apoptotic cell death was determined by DAPI staining and TUNEL assay. Treatment with anti-EpCAM mAb (10 µM) and RAW 264.7 cells for 24 h caused apoptosis; characterized by marked chromatin condensation, small membrane-bound bodies (apoptotic bodies), cytoplasmic condensation, and cellular shrinkage. (A) The TUNEL-positive (green) cells are a apoptotic cells (200× magnification). Apoptotic cells (DAPI-stained TUNEL-positive cells) were estimated by direct counting of fragmented nuclei after DAPI and TUNEL staining. SW620 colon cancer cells (2 × 10 6 /ml) either untreated or treated with doses of anti-EpCAM mAb (10 µM) and RAW264.7 cells. An equal amount of total protein (40 µg/lane) was subjected to 12% SDS-PAGE. (B) Expression of Bax, caspase-3, caspase-8, Bcl-2, TNF-α, and β-actin were detected by western blotting using specific antibodies. Here, β-actin protein was used as an internal control. Each band is representative of 3 independent experiments. Values represent mean (SD) of 3 experiments, each performed in triplicate. * P

    Journal: Experimental & Molecular Medicine

    Article Title: Relationship between ganglioside expression and anti-cancer effects of the monoclonal antibody against epithelial cell adhesion molecule in colon cancer

    doi: 10.3858/emm.2011.43.12.080

    Figure Lengend Snippet: Effects of EpCAM mAb on the induction of apoptosis in colon cancer cells analyzed by DAPI and TUNEL staining. Apoptotic cell death was determined by DAPI staining and TUNEL assay. Treatment with anti-EpCAM mAb (10 µM) and RAW 264.7 cells for 24 h caused apoptosis; characterized by marked chromatin condensation, small membrane-bound bodies (apoptotic bodies), cytoplasmic condensation, and cellular shrinkage. (A) The TUNEL-positive (green) cells are a apoptotic cells (200× magnification). Apoptotic cells (DAPI-stained TUNEL-positive cells) were estimated by direct counting of fragmented nuclei after DAPI and TUNEL staining. SW620 colon cancer cells (2 × 10 6 /ml) either untreated or treated with doses of anti-EpCAM mAb (10 µM) and RAW264.7 cells. An equal amount of total protein (40 µg/lane) was subjected to 12% SDS-PAGE. (B) Expression of Bax, caspase-3, caspase-8, Bcl-2, TNF-α, and β-actin were detected by western blotting using specific antibodies. Here, β-actin protein was used as an internal control. Each band is representative of 3 independent experiments. Values represent mean (SD) of 3 experiments, each performed in triplicate. * P

    Article Snippet: The blots were blocked with 5% non-fat dried milk in Tris-buffered saline for 2 h. The membrane was then incubated for 16 h with a primary antibody: rabbit anti-caspase-8 (Santa Cruz Biotechnology Inc., Delaware, CA), rabbit anti-bax (Santa Cruz Biotechnology Inc.), rabbit anti-caspase-3 (Santa Cruz Biotechnology Inc.), rabbit anti-TNF-α (Santa Cruz Biotechnology Inc.), and mouse monoclonal anti-Bcl-2 (Santa Cruz Biotechnology Inc.), at 4℃.

    Techniques: TUNEL Assay, Staining, SDS Page, Expressing, Western Blot

    Effects of anti-EpCAM mAb on the expression of ganglioside GD1a in apoptosis of colon cancer tumor tissue analyzed by immunohistochemistry and TUNEL. Apoptotic cell death was determined by immunohistochemistry, DAPI staining and TUNEL assay as described in Methods. (A) The TUNEL-positive (green) cells are a apoptotic cells (200× magnification). Apoptotic cells (DAPI-stained, TUNEL-positive cells) were estimated by direct counting of fragmented nuclei after DAPI and TUNEL staining. SW620 colon cancer tumors either untreated or treated with various doses of anti-EpCAM mAb (100 µg). GD1a ganglioside expressing cells were examined with the ABC kit. Anit-EpCAM mAb was injected (100 µg/2 days/mouse) into nude mice for 4 weeks. (B) SW620 cancer tumor tissue as a control (200× magnification). Ganglioside GD1a expression was significantly increased in SW620 cancer tumor tissue (200× magnification). (C) An equal amount of total protein (40 µg/lane) was subjected to 12% SDS-PAGE. Expression of Bax, Caspase-3, Caspase-8, Bcl-2, and β-actin were detected by western blotting using specific antibodies. Here, β-actin protein was used as an internal control. Each band is representative of 3 independent experimental results. The value is the mean (SD) of 3 experiments, with each performed in triplicate. * P

    Journal: Experimental & Molecular Medicine

    Article Title: Relationship between ganglioside expression and anti-cancer effects of the monoclonal antibody against epithelial cell adhesion molecule in colon cancer

    doi: 10.3858/emm.2011.43.12.080

    Figure Lengend Snippet: Effects of anti-EpCAM mAb on the expression of ganglioside GD1a in apoptosis of colon cancer tumor tissue analyzed by immunohistochemistry and TUNEL. Apoptotic cell death was determined by immunohistochemistry, DAPI staining and TUNEL assay as described in Methods. (A) The TUNEL-positive (green) cells are a apoptotic cells (200× magnification). Apoptotic cells (DAPI-stained, TUNEL-positive cells) were estimated by direct counting of fragmented nuclei after DAPI and TUNEL staining. SW620 colon cancer tumors either untreated or treated with various doses of anti-EpCAM mAb (100 µg). GD1a ganglioside expressing cells were examined with the ABC kit. Anit-EpCAM mAb was injected (100 µg/2 days/mouse) into nude mice for 4 weeks. (B) SW620 cancer tumor tissue as a control (200× magnification). Ganglioside GD1a expression was significantly increased in SW620 cancer tumor tissue (200× magnification). (C) An equal amount of total protein (40 µg/lane) was subjected to 12% SDS-PAGE. Expression of Bax, Caspase-3, Caspase-8, Bcl-2, and β-actin were detected by western blotting using specific antibodies. Here, β-actin protein was used as an internal control. Each band is representative of 3 independent experimental results. The value is the mean (SD) of 3 experiments, with each performed in triplicate. * P

    Article Snippet: The blots were blocked with 5% non-fat dried milk in Tris-buffered saline for 2 h. The membrane was then incubated for 16 h with a primary antibody: rabbit anti-caspase-8 (Santa Cruz Biotechnology Inc., Delaware, CA), rabbit anti-bax (Santa Cruz Biotechnology Inc.), rabbit anti-caspase-3 (Santa Cruz Biotechnology Inc.), rabbit anti-TNF-α (Santa Cruz Biotechnology Inc.), and mouse monoclonal anti-Bcl-2 (Santa Cruz Biotechnology Inc.), at 4℃.

    Techniques: Expressing, Immunohistochemistry, TUNEL Assay, Staining, Injection, Mouse Assay, SDS Page, Western Blot

    Human Bcl-2 (hBcl-2) is overexpressed in the odontoblasts of Col2.3Bcl-2 mice and prevents their apoptosis in vivo and in vitro . A. Immunohistochemistry demonstration of hBcl-2 overexpression in the odontoblasts of incisors and molars of 40-day-old Col2.3Bcl-2

    Journal: Journal of cellular biochemistry

    Article Title: Odontoblast-targeted Bcl-2 Overexpression Impairs Dentin Formation

    doi: 10.1002/jcb.22722

    Figure Lengend Snippet: Human Bcl-2 (hBcl-2) is overexpressed in the odontoblasts of Col2.3Bcl-2 mice and prevents their apoptosis in vivo and in vitro . A. Immunohistochemistry demonstration of hBcl-2 overexpression in the odontoblasts of incisors and molars of 40-day-old Col2.3Bcl-2

    Article Snippet: The blots were incubated with 1:100 monoclonal mouse anti-human Bcl-2 (Santa Cruz), mouse anti-mouse Bcl-2 (Santa Cruz), mouse anti-mouse Bax (Santa Cruz), rabbit anti-mouse large fragment (19 kDa) of cleaved caspase-3 (Cell Signaling Technology, Danvers, MA), polyclonal HRP-conjugated goat anti-actin antibodies (Santa Cruz), 1:1000 monoclonal rabbit anti-mouse ERK1/2 (Cell Signaling Technology), or monoclonal rabbit anti-mouse phosphor-ERK1/2 antibody (Cell Signaling Technology) overnight at 4°C.

    Techniques: Mouse Assay, In Vivo, In Vitro, Immunohistochemistry, Over Expression

    Odontoblast-targeted Bcl-2 overexpression results in decreased dentin thickness and mineral density in Col2.3Bcl-2 transgenic mice. A. The top panel shows the cross-sections of incisors where alveolar ridges just appear. The lower panel shows the middle

    Journal: Journal of cellular biochemistry

    Article Title: Odontoblast-targeted Bcl-2 Overexpression Impairs Dentin Formation

    doi: 10.1002/jcb.22722

    Figure Lengend Snippet: Odontoblast-targeted Bcl-2 overexpression results in decreased dentin thickness and mineral density in Col2.3Bcl-2 transgenic mice. A. The top panel shows the cross-sections of incisors where alveolar ridges just appear. The lower panel shows the middle

    Article Snippet: The blots were incubated with 1:100 monoclonal mouse anti-human Bcl-2 (Santa Cruz), mouse anti-mouse Bcl-2 (Santa Cruz), mouse anti-mouse Bax (Santa Cruz), rabbit anti-mouse large fragment (19 kDa) of cleaved caspase-3 (Cell Signaling Technology, Danvers, MA), polyclonal HRP-conjugated goat anti-actin antibodies (Santa Cruz), 1:1000 monoclonal rabbit anti-mouse ERK1/2 (Cell Signaling Technology), or monoclonal rabbit anti-mouse phosphor-ERK1/2 antibody (Cell Signaling Technology) overnight at 4°C.

    Techniques: Over Expression, Transgenic Assay, Mouse Assay

    Bcl-2 overexpression inhibits odontoblast differentiation via a suppression of ERK 1/2 signaling pathway. A. Real-time PCR shows that both wild type and transgenic odontoblasts demonstrate increased expressions of the extracellular matrixes with time,

    Journal: Journal of cellular biochemistry

    Article Title: Odontoblast-targeted Bcl-2 Overexpression Impairs Dentin Formation

    doi: 10.1002/jcb.22722

    Figure Lengend Snippet: Bcl-2 overexpression inhibits odontoblast differentiation via a suppression of ERK 1/2 signaling pathway. A. Real-time PCR shows that both wild type and transgenic odontoblasts demonstrate increased expressions of the extracellular matrixes with time,

    Article Snippet: The blots were incubated with 1:100 monoclonal mouse anti-human Bcl-2 (Santa Cruz), mouse anti-mouse Bcl-2 (Santa Cruz), mouse anti-mouse Bax (Santa Cruz), rabbit anti-mouse large fragment (19 kDa) of cleaved caspase-3 (Cell Signaling Technology, Danvers, MA), polyclonal HRP-conjugated goat anti-actin antibodies (Santa Cruz), 1:1000 monoclonal rabbit anti-mouse ERK1/2 (Cell Signaling Technology), or monoclonal rabbit anti-mouse phosphor-ERK1/2 antibody (Cell Signaling Technology) overnight at 4°C.

    Techniques: Over Expression, Real-time Polymerase Chain Reaction, Transgenic Assay

    Effect of VP16 on the expression and sub-cellular localization of Bax and bcl-2 in ovarian cancer cells. (A) The Western blotting shows that bcl-2 is highly expressed in OVCAR cells and very barely expressed in A2780 cells. VP16 greatly reduces (∼50%) the cellular content of bcl-2 in OVCAR cells, whereas it does not affect the expression of this protein in A2780 cells (see densitometry). (B) The Western blotting shows that Bax is highly expressed in A2780 and faintly detectable in OVCAR. Bax expression is not affected by VP16 (see densitometry). (C) Immnunolocalization of bcl-2 and Bax. Both proteins appear as barely detectable spots diffused in the cytoplasm in control cells. On treatment with VP16 only in A2780 cells Bax appears as strongly intense spots in confined region of the cytoplasm, indicative of conformational activation and granular-like aggregation of the molecules. Bcl-2 does not co-localize with such Bax-positive aggregates.

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Chemotherapy drug response in ovarian cancer cells strictly depends on a cathepsin D-Bax activation loop

    doi: 10.1111/j.1582-4934.2008.00435.x

    Figure Lengend Snippet: Effect of VP16 on the expression and sub-cellular localization of Bax and bcl-2 in ovarian cancer cells. (A) The Western blotting shows that bcl-2 is highly expressed in OVCAR cells and very barely expressed in A2780 cells. VP16 greatly reduces (∼50%) the cellular content of bcl-2 in OVCAR cells, whereas it does not affect the expression of this protein in A2780 cells (see densitometry). (B) The Western blotting shows that Bax is highly expressed in A2780 and faintly detectable in OVCAR. Bax expression is not affected by VP16 (see densitometry). (C) Immnunolocalization of bcl-2 and Bax. Both proteins appear as barely detectable spots diffused in the cytoplasm in control cells. On treatment with VP16 only in A2780 cells Bax appears as strongly intense spots in confined region of the cytoplasm, indicative of conformational activation and granular-like aggregation of the molecules. Bcl-2 does not co-localize with such Bax-positive aggregates.

    Article Snippet: The filter was probed with specific monoclonal antibodies against CD (monoclonal from EMD Biosciences, Calbiochem, San Diego, CA, USA), or mouse monoclonal antibody bcl-2 (Santa Cruz Technology, Inc., Santa Cruz, CA, USA) or rabbit polyclonal anti-human Bax antiserum (Cell Signaling Technology, MA, USA) or mouse monoclonal anti-human p53 (DO-1 Santa Cruz Biotechnology, Inc.) or mouse monoclonal α-actin (Sigma-Aldrich Corp.).

    Techniques: Expressing, Western Blot, Activation Assay

    Mitochondrial-BCL-2 preferentially binds AMBRA1, and this binding is disrupted after autophagy induction. ( A ) HEK293 cells were co-transfected with vectors encoding either ER-BCL-2 or mito-BCL-2 and myc-AMBRA1. Cells were grown either in normal media

    Journal: The EMBO Journal

    Article Title: Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy

    doi: 10.1038/emboj.2011.49

    Figure Lengend Snippet: Mitochondrial-BCL-2 preferentially binds AMBRA1, and this binding is disrupted after autophagy induction. ( A ) HEK293 cells were co-transfected with vectors encoding either ER-BCL-2 or mito-BCL-2 and myc-AMBRA1. Cells were grown either in normal media

    Article Snippet: After rinsing in medium containing 0.01% Tween-20 (Merck), a second incubation with primary antibody mouse anti-BCL-2 (Santa Cruz) diluted 1:50 in medium A was performed, for 2 h at RT.

    Techniques: Binding Assay, Transfection

    AMBRA1 does not posses a proapoptotic activity but, in contrast, it may participate to cell viability. ( A ) AMBRA1–BCL-2 interaction is disrupted after stautosporine (STS) treatment. HEK293 cells were co-transfected with vectors encoding mito-BCL-2

    Journal: The EMBO Journal

    Article Title: Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy

    doi: 10.1038/emboj.2011.49

    Figure Lengend Snippet: AMBRA1 does not posses a proapoptotic activity but, in contrast, it may participate to cell viability. ( A ) AMBRA1–BCL-2 interaction is disrupted after stautosporine (STS) treatment. HEK293 cells were co-transfected with vectors encoding mito-BCL-2

    Article Snippet: After rinsing in medium containing 0.01% Tween-20 (Merck), a second incubation with primary antibody mouse anti-BCL-2 (Santa Cruz) diluted 1:50 in medium A was performed, for 2 h at RT.

    Techniques: Activity Assay, Transfection

    The interaction between AMBRA1 and BCL-2 does not require BECLIN 1. ( A ). ( B ) BECLIN

    Journal: The EMBO Journal

    Article Title: Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy

    doi: 10.1038/emboj.2011.49

    Figure Lengend Snippet: The interaction between AMBRA1 and BCL-2 does not require BECLIN 1. ( A ). ( B ) BECLIN

    Article Snippet: After rinsing in medium containing 0.01% Tween-20 (Merck), a second incubation with primary antibody mouse anti-BCL-2 (Santa Cruz) diluted 1:50 in medium A was performed, for 2 h at RT.

    Techniques:

    AMBRA1 dynamic colocalization with mito-BCL-2 and the mitochondrial network. ( A , B ) HEK293 cells co-transfected with vectors encoding mito-BCL-2 and myc-AMBRA1, grown either in normal media ( A ) or in EBSS ( B ) for 4 h and stained with an anti-BCL-2 antibody

    Journal: The EMBO Journal

    Article Title: Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy

    doi: 10.1038/emboj.2011.49

    Figure Lengend Snippet: AMBRA1 dynamic colocalization with mito-BCL-2 and the mitochondrial network. ( A , B ) HEK293 cells co-transfected with vectors encoding mito-BCL-2 and myc-AMBRA1, grown either in normal media ( A ) or in EBSS ( B ) for 4 h and stained with an anti-BCL-2 antibody

    Article Snippet: After rinsing in medium containing 0.01% Tween-20 (Merck), a second incubation with primary antibody mouse anti-BCL-2 (Santa Cruz) diluted 1:50 in medium A was performed, for 2 h at RT.

    Techniques: Transfection, Staining

    AMBRA1 proautophagic activity is inhibited by mito-BCL-2. ( A ) HEK293 cells were co-transfected either with the control vector pcDNA3, vectors encoding Mito-, ER- and WT-BCL-2 as controls and a vector encoding myc–AMBRA1 or AMBRA1-F2 with or without

    Journal: The EMBO Journal

    Article Title: Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy

    doi: 10.1038/emboj.2011.49

    Figure Lengend Snippet: AMBRA1 proautophagic activity is inhibited by mito-BCL-2. ( A ) HEK293 cells were co-transfected either with the control vector pcDNA3, vectors encoding Mito-, ER- and WT-BCL-2 as controls and a vector encoding myc–AMBRA1 or AMBRA1-F2 with or without

    Article Snippet: After rinsing in medium containing 0.01% Tween-20 (Merck), a second incubation with primary antibody mouse anti-BCL-2 (Santa Cruz) diluted 1:50 in medium A was performed, for 2 h at RT.

    Techniques: Activity Assay, Transfection, Plasmid Preparation

    AMBRA1 interacts with the antiapoptotic factor BCL-2 in mammalian cells. A scheme of AMBRA1 with its WD40 domains is illustrated in ( A ). The binding site with BECLIN 1 is also reported on AMBRA1. HEK293 cells were co-transfected with vectors encoding

    Journal: The EMBO Journal

    Article Title: Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy

    doi: 10.1038/emboj.2011.49

    Figure Lengend Snippet: AMBRA1 interacts with the antiapoptotic factor BCL-2 in mammalian cells. A scheme of AMBRA1 with its WD40 domains is illustrated in ( A ). The binding site with BECLIN 1 is also reported on AMBRA1. HEK293 cells were co-transfected with vectors encoding

    Article Snippet: After rinsing in medium containing 0.01% Tween-20 (Merck), a second incubation with primary antibody mouse anti-BCL-2 (Santa Cruz) diluted 1:50 in medium A was performed, for 2 h at RT.

    Techniques: Binding Assay, Transfection

    AMBRA1 competes with BCL-2 to bind BECLIN 1. ( A ) Illustration of the BECLIN 1 mutants. The coiled-coiled domain (CCD) and the evolutionary conserved domain (ECD) of BECLIN 1 are indicated. ( B ) AMBRA1 binds all truncated mutants of BECLIN 1. Co-immunoprecipitation

    Journal: The EMBO Journal

    Article Title: Mitochondrial BCL-2 inhibits AMBRA1-induced autophagy

    doi: 10.1038/emboj.2011.49

    Figure Lengend Snippet: AMBRA1 competes with BCL-2 to bind BECLIN 1. ( A ) Illustration of the BECLIN 1 mutants. The coiled-coiled domain (CCD) and the evolutionary conserved domain (ECD) of BECLIN 1 are indicated. ( B ) AMBRA1 binds all truncated mutants of BECLIN 1. Co-immunoprecipitation

    Article Snippet: After rinsing in medium containing 0.01% Tween-20 (Merck), a second incubation with primary antibody mouse anti-BCL-2 (Santa Cruz) diluted 1:50 in medium A was performed, for 2 h at RT.

    Techniques: Immunoprecipitation

    Only enzymatically active FKBP38 interacts with Bcl-2. ( A ) Co-immunoprecipitation of endogenous FKBP38 and Bcl-2. After preincubation with 500 μM EGTA, SH-SY5Y cell lysate was incubated with rabbit anti-FKBP38 antibody. Antibody/protein complexes were bound to protein G-Sepharose. Samples were washed. Precipitates (lanes 1–3) and input were subjected to SDS–PAGE and analyzed by Western blot using mouse anti-Bcl-2 antibody. Cell lysate preincubated with rabbit immunoglobin was used as a control. FKBP38 bound to Bcl-2 in the presence of 1 mM calcium. This interaction was disrupted by 1 μM GPI1046. No interaction was observed in the absence of Ca 2+ . ( B ) SH-SY5Y crude cell extract was applied in the absence and presence of 20 μM CaM and 2 mM Ca 2+ to maltose-binding protein-Bcl-2 fusion protein (MBP-Bcl-2) immobilized on amylose resin. After three washing steps, protein was eluted by 200 mM maltose and analyzed by Western blot using polyclonal anti-FKBP38 antibody. The proportion of active endogenous FKBP38 was quantified by Biorad Multi-Analyst software. ( C ) MBP-Bcl-2 was immobilized on amylose resin and incubated with (1) FKBP38, (2) FKBP38 and CaM, (3) FKBP38 and Ca 2+ /CaM and (4) FKBP38, Ca 2+ /CaM and 200 nM GPI1046. After washing, protein was eluted and analyzed by Western blot using polyclonal anti-FKBP38 antibody. ( D ) Inhibition of PPIase activity of 1 μM FKBP38 by Bcl-2 was measured in the PPIase assay in the presence of 5 mM CaCl 2 and 5 μM CaM. The calculated K i value was 0.74 μM.

    Journal: The EMBO Journal

    Article Title: Bcl-2 regulator FKBP38 is activated by Ca2+/calmodulin

    doi: 10.1038/sj.emboj.7600739

    Figure Lengend Snippet: Only enzymatically active FKBP38 interacts with Bcl-2. ( A ) Co-immunoprecipitation of endogenous FKBP38 and Bcl-2. After preincubation with 500 μM EGTA, SH-SY5Y cell lysate was incubated with rabbit anti-FKBP38 antibody. Antibody/protein complexes were bound to protein G-Sepharose. Samples were washed. Precipitates (lanes 1–3) and input were subjected to SDS–PAGE and analyzed by Western blot using mouse anti-Bcl-2 antibody. Cell lysate preincubated with rabbit immunoglobin was used as a control. FKBP38 bound to Bcl-2 in the presence of 1 mM calcium. This interaction was disrupted by 1 μM GPI1046. No interaction was observed in the absence of Ca 2+ . ( B ) SH-SY5Y crude cell extract was applied in the absence and presence of 20 μM CaM and 2 mM Ca 2+ to maltose-binding protein-Bcl-2 fusion protein (MBP-Bcl-2) immobilized on amylose resin. After three washing steps, protein was eluted by 200 mM maltose and analyzed by Western blot using polyclonal anti-FKBP38 antibody. The proportion of active endogenous FKBP38 was quantified by Biorad Multi-Analyst software. ( C ) MBP-Bcl-2 was immobilized on amylose resin and incubated with (1) FKBP38, (2) FKBP38 and CaM, (3) FKBP38 and Ca 2+ /CaM and (4) FKBP38, Ca 2+ /CaM and 200 nM GPI1046. After washing, protein was eluted and analyzed by Western blot using polyclonal anti-FKBP38 antibody. ( D ) Inhibition of PPIase activity of 1 μM FKBP38 by Bcl-2 was measured in the PPIase assay in the presence of 5 mM CaCl 2 and 5 μM CaM. The calculated K i value was 0.74 μM.

    Article Snippet: Additional antibodies used were polyclonal rabbit anti-actin (Sigma), monoclonal hamster anti-Bcl-2 (Pharmingen), monoclonal mouse anti-Bcl-2 (Santa Cruz), polyclonal rabbit anti-CaM (Santa Cruz), monoclonal mouse anti-CaM (Sigma), monoclonal mouse anti-Bad (Pharmingen) and polyclonal goat anti-mouse ImageBlue labeled (Leinco).

    Techniques: Immunoprecipitation, Incubation, SDS Page, Western Blot, Chick Chorioallantoic Membrane Assay, Binding Assay, Software, Inhibition, Activity Assay

    ( C ) Subcellular localization of FKBP38 and Bcl-2 was studied by preparing mitochondria, ER, nucleus and cytosol from SH-SY5Y cells and analyzing these fractions by Western blotting using anti-FKBP38 and anti-Bcl-2 antibodies. Antibodies detecting cytochrome c (Cyt c ) and FKBP13 were used as controls for mitochondrial and ER localization, respectively.

    Journal: The EMBO Journal

    Article Title: Bcl-2 regulator FKBP38 is activated by Ca2+/calmodulin

    doi: 10.1038/sj.emboj.7600739

    Figure Lengend Snippet: ( C ) Subcellular localization of FKBP38 and Bcl-2 was studied by preparing mitochondria, ER, nucleus and cytosol from SH-SY5Y cells and analyzing these fractions by Western blotting using anti-FKBP38 and anti-Bcl-2 antibodies. Antibodies detecting cytochrome c (Cyt c ) and FKBP13 were used as controls for mitochondrial and ER localization, respectively.

    Article Snippet: Additional antibodies used were polyclonal rabbit anti-actin (Sigma), monoclonal hamster anti-Bcl-2 (Pharmingen), monoclonal mouse anti-Bcl-2 (Santa Cruz), polyclonal rabbit anti-CaM (Santa Cruz), monoclonal mouse anti-CaM (Sigma), monoclonal mouse anti-Bad (Pharmingen) and polyclonal goat anti-mouse ImageBlue labeled (Leinco).

    Techniques: Western Blot

    FKBP38 and Bcl-2 colocalize in neuroblastoma cells. ( A ) Localization of Bcl-2 and FKBP38 in SH-SY5Y neuroblastoma cells was analyzed by immunostaining with Cy5-conjugated goat anti-hamster antibody against hamster anti-Bcl-2 antibody and FITC-conjugated goat anti-rabbit IgG against rabbit anti-FKBP38 antibody. Cells were treated for 16 h with 50 μM etoposide and 2 μM GPI1046. Nuclei were stained with DAPI. ( B ) Subcellular distribution of FKBP38 in SH-SY5Y cells was studied by immunostaining with FITC-conjugated goat anti-rabbit IgG against rabbit anti-FKBP38 antibody and a subcellular structure localization kit (Chemicon).

    Journal: The EMBO Journal

    Article Title: Bcl-2 regulator FKBP38 is activated by Ca2+/calmodulin

    doi: 10.1038/sj.emboj.7600739

    Figure Lengend Snippet: FKBP38 and Bcl-2 colocalize in neuroblastoma cells. ( A ) Localization of Bcl-2 and FKBP38 in SH-SY5Y neuroblastoma cells was analyzed by immunostaining with Cy5-conjugated goat anti-hamster antibody against hamster anti-Bcl-2 antibody and FITC-conjugated goat anti-rabbit IgG against rabbit anti-FKBP38 antibody. Cells were treated for 16 h with 50 μM etoposide and 2 μM GPI1046. Nuclei were stained with DAPI. ( B ) Subcellular distribution of FKBP38 in SH-SY5Y cells was studied by immunostaining with FITC-conjugated goat anti-rabbit IgG against rabbit anti-FKBP38 antibody and a subcellular structure localization kit (Chemicon).

    Article Snippet: Additional antibodies used were polyclonal rabbit anti-actin (Sigma), monoclonal hamster anti-Bcl-2 (Pharmingen), monoclonal mouse anti-Bcl-2 (Santa Cruz), polyclonal rabbit anti-CaM (Santa Cruz), monoclonal mouse anti-CaM (Sigma), monoclonal mouse anti-Bad (Pharmingen) and polyclonal goat anti-mouse ImageBlue labeled (Leinco).

    Techniques: Immunostaining, Staining

    FKBP38 activity influences Bcl-2 function. ( A ) Near-UV CD spectra of 1 μM Ca 2+ /CaM/FKBP38 and 3 μM Bcl-2 were measured either separated (dotted line) or mixed (solid line) in a tandem cuvette. ( B ) Co-immunoprecipitation. SH-SY5Y cells were stimulated to apoptosis by 50 μM etoposide. Cell lysate preincubated with 500 μM EGTA was incubated with monoclonal mouse anti-Bcl-2 antibody. Antibody/protein complexes were bound to protein G-Sepharose. Samples were washed. Precipitates (lanes 1–3) and input were subjected to SDS–PAGE and analyzed by Western blot using rabbit anti-FKBP38 antibody. Cell lysate preincubated with control mouse immunoglobin was used as a control. FKBP38 bound to Bcl-2 in the presence of 1 mM Ca 2+ , whereas 1 μM GPI1046 abolished the interaction. Bcl-2/Bad interaction was investigated by incubation of SH-SY5Y cell lysate with hamster anti-Bcl-2 antibody and protein was detected using mouse anti-Bad antibody. Bad/Bcl-2 interaction was observed in the presence of 1 mM calcium and 1 μM GPI1046. In addition, SH-SY5Y cell lysate was incubated with monoclonal mouse anti-Bad antibody, and antibody/protein complexes formed were bound to protein G-Sepharose. Precipitates (lanes 1–3) and input were subjected to SDS–PAGE and analyzed by Western blot using hamster anti-Bcl-2 antibody. Cell lysate preincubated with mouse immunoglobin was used as a control. Bcl-2 bound to Bad in the presence of 1 mM Ca 2+ and 1 μM GPI1046, indicating that GPI1046 interfered with the Bcl-2/FKBP38 interaction, allowing Bad/Bcl-2 complexes to form. ( C ) MBP-Bcl-2 fusion protein was immobilized on amylose beads and incubated with Bad (Sigma), FKBP38 and CaM either in the presence or absence of 2 mM Ca 2+ . Pellet (P) and supernatant (S) were subjected to 12.5% SDS–PAGE and analyzed by Western blotting using mouse anti-Bad antibody and hamster anti-Bcl-2 antibody. ( D ) Subcellular distribution of Bcl-2 and Bad in SH-SY5Y neuroblastoma cells was analyzed by immunostaining with FITC-conjugated goat anti-mouse antibody against mouse anti-Bad antibody and Cy5-conjugated goat anti-hamster antibody against hamster anti-Bcl-2 antibody. Nuclei were stained with DAPI. Cells were treated for 16 h with 50 μM etoposide either in the absence or presence of 1 μM GPI1046. Cells transfected with FKBP38 RNAi construct were analyzed with ImageBlue-conjugated goat anti-mouse antibody against mouse anti-Bad antibody and TRITC-conjugated goat anti-hamster antibody against hamster anti-Bcl-2. Nuclei were stained by 7-AAD. Because of better understanding, colors of this panel were adapted to other results presented in this figure. Again, inhibition of FKBP38 activity allows Bcl-2/Bad interaction after induction of apoptosis by 50 μM etoposide.

    Journal: The EMBO Journal

    Article Title: Bcl-2 regulator FKBP38 is activated by Ca2+/calmodulin

    doi: 10.1038/sj.emboj.7600739

    Figure Lengend Snippet: FKBP38 activity influences Bcl-2 function. ( A ) Near-UV CD spectra of 1 μM Ca 2+ /CaM/FKBP38 and 3 μM Bcl-2 were measured either separated (dotted line) or mixed (solid line) in a tandem cuvette. ( B ) Co-immunoprecipitation. SH-SY5Y cells were stimulated to apoptosis by 50 μM etoposide. Cell lysate preincubated with 500 μM EGTA was incubated with monoclonal mouse anti-Bcl-2 antibody. Antibody/protein complexes were bound to protein G-Sepharose. Samples were washed. Precipitates (lanes 1–3) and input were subjected to SDS–PAGE and analyzed by Western blot using rabbit anti-FKBP38 antibody. Cell lysate preincubated with control mouse immunoglobin was used as a control. FKBP38 bound to Bcl-2 in the presence of 1 mM Ca 2+ , whereas 1 μM GPI1046 abolished the interaction. Bcl-2/Bad interaction was investigated by incubation of SH-SY5Y cell lysate with hamster anti-Bcl-2 antibody and protein was detected using mouse anti-Bad antibody. Bad/Bcl-2 interaction was observed in the presence of 1 mM calcium and 1 μM GPI1046. In addition, SH-SY5Y cell lysate was incubated with monoclonal mouse anti-Bad antibody, and antibody/protein complexes formed were bound to protein G-Sepharose. Precipitates (lanes 1–3) and input were subjected to SDS–PAGE and analyzed by Western blot using hamster anti-Bcl-2 antibody. Cell lysate preincubated with mouse immunoglobin was used as a control. Bcl-2 bound to Bad in the presence of 1 mM Ca 2+ and 1 μM GPI1046, indicating that GPI1046 interfered with the Bcl-2/FKBP38 interaction, allowing Bad/Bcl-2 complexes to form. ( C ) MBP-Bcl-2 fusion protein was immobilized on amylose beads and incubated with Bad (Sigma), FKBP38 and CaM either in the presence or absence of 2 mM Ca 2+ . Pellet (P) and supernatant (S) were subjected to 12.5% SDS–PAGE and analyzed by Western blotting using mouse anti-Bad antibody and hamster anti-Bcl-2 antibody. ( D ) Subcellular distribution of Bcl-2 and Bad in SH-SY5Y neuroblastoma cells was analyzed by immunostaining with FITC-conjugated goat anti-mouse antibody against mouse anti-Bad antibody and Cy5-conjugated goat anti-hamster antibody against hamster anti-Bcl-2 antibody. Nuclei were stained with DAPI. Cells were treated for 16 h with 50 μM etoposide either in the absence or presence of 1 μM GPI1046. Cells transfected with FKBP38 RNAi construct were analyzed with ImageBlue-conjugated goat anti-mouse antibody against mouse anti-Bad antibody and TRITC-conjugated goat anti-hamster antibody against hamster anti-Bcl-2. Nuclei were stained by 7-AAD. Because of better understanding, colors of this panel were adapted to other results presented in this figure. Again, inhibition of FKBP38 activity allows Bcl-2/Bad interaction after induction of apoptosis by 50 μM etoposide.

    Article Snippet: Additional antibodies used were polyclonal rabbit anti-actin (Sigma), monoclonal hamster anti-Bcl-2 (Pharmingen), monoclonal mouse anti-Bcl-2 (Santa Cruz), polyclonal rabbit anti-CaM (Santa Cruz), monoclonal mouse anti-CaM (Sigma), monoclonal mouse anti-Bad (Pharmingen) and polyclonal goat anti-mouse ImageBlue labeled (Leinco).

    Techniques: Activity Assay, Chick Chorioallantoic Membrane Assay, Immunoprecipitation, Incubation, SDS Page, Western Blot, Immunostaining, Staining, Transfection, Construct, Inhibition

    Semiquantitative RT–PCR and Western blotting revealed that mRNA ( a ) and protein ( b ) levels of Bcl-2, an anti-apoptotic factor, in both normal thymuses and thymic tumors of heterozygous (+/−) and homozygous (−/−) mice were significantly higher than in wild-type (+/+) siblings. n =6; a p

    Journal: Hormones & cancer

    Article Title: Luteinizing Hormone Receptor Deficiency Increases the Susceptibility to Alkylating Agent-Induced Lymphomagenesis in Mice

    doi: 10.1007/s12672-010-0045-3

    Figure Lengend Snippet: Semiquantitative RT–PCR and Western blotting revealed that mRNA ( a ) and protein ( b ) levels of Bcl-2, an anti-apoptotic factor, in both normal thymuses and thymic tumors of heterozygous (+/−) and homozygous (−/−) mice were significantly higher than in wild-type (+/+) siblings. n =6; a p

    Article Snippet: After blocking with 3% nonfat dry milk, the membranes were incubated overnight at 4°C with 1:1,000 diluted rabbit polyclonal antibodies against un-cleaved (inactive) and cleaved (active) caspase-3 (9662 and 9661; Cell Signaling, Beverly, MA, USA), 1:500 diluted mouse monoclonal antibody against Bcl-2 (sc-509; Santa Cruz Biotech), and 1:1,000 diluted mouse monoclonal antibody against β-actin (A5441; Sigma), respectively.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Mouse Assay

    Bcl-2 inhibitor enhances the effect of psoralidin on SW480 cells. (A) Effect of Bcl-2 inhibitor on Bcl-2 protein expression of SW480 cells by western blotting assays; (B) statistical analysis of Bcl-2 protein expression of SW480 cells; and (C) Bcl-2 inhibitor enhances the effect of psoralidin on viability of SW480 cells. Values are expressed as the mean ± standard deviation. **P

    Journal: Oncology Letters

    Article Title: Differential effect of psoralidin in enhancing apoptosis of colon cancer cells via nuclear factor-κB and B-cell lymphoma-2/B-cell lymphoma-2-associated X protein signaling pathways

    doi: 10.3892/ol.2015.3861

    Figure Lengend Snippet: Bcl-2 inhibitor enhances the effect of psoralidin on SW480 cells. (A) Effect of Bcl-2 inhibitor on Bcl-2 protein expression of SW480 cells by western blotting assays; (B) statistical analysis of Bcl-2 protein expression of SW480 cells; and (C) Bcl-2 inhibitor enhances the effect of psoralidin on viability of SW480 cells. Values are expressed as the mean ± standard deviation. **P

    Article Snippet: Following blocking with Tris-buffered saline (Beyotime Institute of Biotechnology) containing 5% non-fat milk for 2 h, the membranes were incubated with monoclonal mouse anti-human Bcl-2 (1:1,500; cat. no. sc-7382; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), monoclonal mouse anti-human Bax (1:2,000; cat. no. sc-8432; Santa Cruz Biotechnology, Inc.) and monoclonal mouse anti-human β-actin (1:500; cat. no. sc-8432; Tiangen Biotech Co., Ltd., Beijing, China) overnight at 4°C.

    Techniques: Expressing, Western Blot, Standard Deviation

    Psoralidin inhibits Bcl-2 and enhances Bax protein expression in SW480 cells. (A and B) Effect of psoralidin on Bcl-2 protein expression of SW480 cells by western blotting assay and statistical analysis of Bcl-2 protein expression of SW480 cells. (C and D) Effects of psoralidin on Bax protein expression of SW480 cells by western blotting assay and statistical analysis of Bax proteins expression of SW480 cells. Values are expressed as the mean ± standard deviation. *P

    Journal: Oncology Letters

    Article Title: Differential effect of psoralidin in enhancing apoptosis of colon cancer cells via nuclear factor-κB and B-cell lymphoma-2/B-cell lymphoma-2-associated X protein signaling pathways

    doi: 10.3892/ol.2015.3861

    Figure Lengend Snippet: Psoralidin inhibits Bcl-2 and enhances Bax protein expression in SW480 cells. (A and B) Effect of psoralidin on Bcl-2 protein expression of SW480 cells by western blotting assay and statistical analysis of Bcl-2 protein expression of SW480 cells. (C and D) Effects of psoralidin on Bax protein expression of SW480 cells by western blotting assay and statistical analysis of Bax proteins expression of SW480 cells. Values are expressed as the mean ± standard deviation. *P

    Article Snippet: Following blocking with Tris-buffered saline (Beyotime Institute of Biotechnology) containing 5% non-fat milk for 2 h, the membranes were incubated with monoclonal mouse anti-human Bcl-2 (1:1,500; cat. no. sc-7382; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), monoclonal mouse anti-human Bax (1:2,000; cat. no. sc-8432; Santa Cruz Biotechnology, Inc.) and monoclonal mouse anti-human β-actin (1:500; cat. no. sc-8432; Tiangen Biotech Co., Ltd., Beijing, China) overnight at 4°C.

    Techniques: Expressing, Western Blot, Standard Deviation

    Regulation of intracerebral gene expression for apoptotic mediators . Total RNA was extracted from homogenized murine brains (injured left hemispheres) at 4 h, 24 h, and 7 days after sham surgery or experimental head injury. Gene expression levels of Fas, FasL, Bax and Bcl-2 were determined by semi-quantitative two-step real-time RT-PCR, as described in the methods section. A significant induction of Fas gene expression was detected at 4 h in both trauma groups, compared to sham-operated controls (* P

    Journal: Journal of Neuroinflammation

    Article Title: Absence of the complement regulatory molecule CD59a leads to exacerbated neuropathology after traumatic brain injury in mice

    doi: 10.1186/1742-2094-6-2

    Figure Lengend Snippet: Regulation of intracerebral gene expression for apoptotic mediators . Total RNA was extracted from homogenized murine brains (injured left hemispheres) at 4 h, 24 h, and 7 days after sham surgery or experimental head injury. Gene expression levels of Fas, FasL, Bax and Bcl-2 were determined by semi-quantitative two-step real-time RT-PCR, as described in the methods section. A significant induction of Fas gene expression was detected at 4 h in both trauma groups, compared to sham-operated controls (* P

    Article Snippet: The blots were blocked overnight and then incubated with either polyclonal rabbit anti-mouse Fas (1:200), rabbit anti-mouse FasL (1:200), rabbit anti-mouse Bax (1:300), or monoclonal anti-mouse Bcl-2 (1:500) antibodies (Santa Cruz Biotechnology, Heidelberg, Germany), and with a monocloncal anti-β-actin antibody (clone AC-15, Sigma) diluted 1:10,000, as internal control for ascertaining equal loading of the bands.

    Techniques: Expressing, Quantitative RT-PCR

    Gene expression of apoptosis regulatory proteins. Effects of CXB (20 μM) on mRNA expression of caspase-3 ( a ), Bax ( b ), Bcl-2 ( c ), and the Bcl-2/Bax ratio ( d ). * P

    Journal: Molecular Biology Reports

    Article Title: Celecoxib inhibits growth of human autosomal dominant polycystic kidney cyst-lining epithelial cells through the VEGF/Raf/MAPK/ERK signaling pathway

    doi: 10.1007/s11033-012-1611-2

    Figure Lengend Snippet: Gene expression of apoptosis regulatory proteins. Effects of CXB (20 μM) on mRNA expression of caspase-3 ( a ), Bax ( b ), Bcl-2 ( c ), and the Bcl-2/Bax ratio ( d ). * P

    Article Snippet: Protein (40 μg) from each dish was loaded for polyacrylamide gel electrophoresis (SDS-PAGE, resolving gel concentration was 12%) and transferred to nitrocellulose membranes at 100 V for 2 h. The membrane was blocked by treatment with 5% nonfat milk for 1 h at room temperature and was probed at 4°C overnight with 11 different primary antibodies: mouse anti-human PCNA (1:200, Santa Cruz), mouse anti-human ERK1/2 (1:2,000, Cell Signaling Technology), rabbit anti-human phospho-ERK1/2 (1:1,000, Cell Signaling Technology), mouse anti-human p21CIP/WAF1 (1:200, Santa Cruz), mouse anti-human Cyclin A (1:1,000), mouse anti-human Bax (1:100, Santa Cruz), mouse anti-human Bcl-2 (1:100, Santa Cruz), mouse anti-human caspase-3 (1:800, Santa Cruz), rabbit anti-human VEGFR-2 (1:800, Santa Cruz), rabbit anti-human Raf-1 (1:800, Santa Cruz) and mouse anti-human GAPDH (Sigma).

    Techniques: Expressing

    Protein expression of cell cycle and apoptosis regulatory proteins. Effects of CXB (20 μM) on ERK1/2 signaling pathway phosphorylation ( a ), PCNA expression ( a ), Bax/Bcl-2 expression ( b ), caspase-3 cleavage ( b ), Cyclin A expression ( b ) and p21 CIP/WAF1 expression ( b ). GAPDH was used as a loading control. * P

    Journal: Molecular Biology Reports

    Article Title: Celecoxib inhibits growth of human autosomal dominant polycystic kidney cyst-lining epithelial cells through the VEGF/Raf/MAPK/ERK signaling pathway

    doi: 10.1007/s11033-012-1611-2

    Figure Lengend Snippet: Protein expression of cell cycle and apoptosis regulatory proteins. Effects of CXB (20 μM) on ERK1/2 signaling pathway phosphorylation ( a ), PCNA expression ( a ), Bax/Bcl-2 expression ( b ), caspase-3 cleavage ( b ), Cyclin A expression ( b ) and p21 CIP/WAF1 expression ( b ). GAPDH was used as a loading control. * P

    Article Snippet: Protein (40 μg) from each dish was loaded for polyacrylamide gel electrophoresis (SDS-PAGE, resolving gel concentration was 12%) and transferred to nitrocellulose membranes at 100 V for 2 h. The membrane was blocked by treatment with 5% nonfat milk for 1 h at room temperature and was probed at 4°C overnight with 11 different primary antibodies: mouse anti-human PCNA (1:200, Santa Cruz), mouse anti-human ERK1/2 (1:2,000, Cell Signaling Technology), rabbit anti-human phospho-ERK1/2 (1:1,000, Cell Signaling Technology), mouse anti-human p21CIP/WAF1 (1:200, Santa Cruz), mouse anti-human Cyclin A (1:1,000), mouse anti-human Bax (1:100, Santa Cruz), mouse anti-human Bcl-2 (1:100, Santa Cruz), mouse anti-human caspase-3 (1:800, Santa Cruz), rabbit anti-human VEGFR-2 (1:800, Santa Cruz), rabbit anti-human Raf-1 (1:800, Santa Cruz) and mouse anti-human GAPDH (Sigma).

    Techniques: Expressing

    Ursolic acid regulates the expression of apoptosis-associated proteins in SNU-484 cells. Cells were treated with various concentrations of ursolic acid (0, 1.0, 2.5 and 5.0 μM) for 24 h and the protein expression levels of levels of (A) PARP, (B) pro-caspase 3, (C) Bcl-2 and (D) Bax were determined by performing an immunoblot analysis. * P

    Journal: Oncology Letters

    Article Title: Ursolic acid inhibits the invasive phenotype of SNU-484 human gastric cancer cells

    doi: 10.3892/ol.2014.2735

    Figure Lengend Snippet: Ursolic acid regulates the expression of apoptosis-associated proteins in SNU-484 cells. Cells were treated with various concentrations of ursolic acid (0, 1.0, 2.5 and 5.0 μM) for 24 h and the protein expression levels of levels of (A) PARP, (B) pro-caspase 3, (C) Bcl-2 and (D) Bax were determined by performing an immunoblot analysis. * P

    Article Snippet: Mouse monoclonal anti-human Bcl-2, mouse monoclonal anti-human caspase-3 and rabbit polyclonal anti-human Bax antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

    Techniques: Expressing

    Protein levels of Bcl-2/Bim in CD4 + CD25 − T cells after CLP induced sepsis. (A) Using Western blot, Bcl-2 and Bim protein expressions were determined both in vivo and in vitro . (B) Expression of antiapoptotic protein Bcl-2 in CD4 + CD25 − T cells was significantly decreased after septic challenge ( P

    Journal: Journal of Interferon & Cytokine Research

    Article Title: Effect of Regulatory T Cells on Promoting Apoptosis of T Lymphocyte and Its Regulatory Mechanism in Sepsis

    doi: 10.1089/jir.2014.0235

    Figure Lengend Snippet: Protein levels of Bcl-2/Bim in CD4 + CD25 − T cells after CLP induced sepsis. (A) Using Western blot, Bcl-2 and Bim protein expressions were determined both in vivo and in vitro . (B) Expression of antiapoptotic protein Bcl-2 in CD4 + CD25 − T cells was significantly decreased after septic challenge ( P

    Article Snippet: Rat anti-mouse Bcl-2 and the Bim antibody were purchased from Santa Cruz Biotechnology.

    Techniques: Western Blot, In Vivo, In Vitro, Expressing

    The protein expression of Smad2/Smad and P-Smad2/P-Smad3 in the CD4 + CD25 − T cells after polymicrobial sepsis. (A) Levels of Smad2/Smad3, P-Smad2/Smad3, and Bcl-2 superfamily members of Bcl-2/Bim in CD4 + CD25 − T cells were determined by Western blot analysis. (B) There were no significant differences of Smad2/Smad3 expression in CD4 + CD25 − T cells among various groups; the expressions of P-smad2/P-Smad3 in the CLP group were significantly higher than that in sham group, while P-smad2/P-Smad3 expression in CLP+(PC61 or anti-TGF-β antibody) group was significantly lower than that in the CLP group ( P

    Journal: Journal of Interferon & Cytokine Research

    Article Title: Effect of Regulatory T Cells on Promoting Apoptosis of T Lymphocyte and Its Regulatory Mechanism in Sepsis

    doi: 10.1089/jir.2014.0235

    Figure Lengend Snippet: The protein expression of Smad2/Smad and P-Smad2/P-Smad3 in the CD4 + CD25 − T cells after polymicrobial sepsis. (A) Levels of Smad2/Smad3, P-Smad2/Smad3, and Bcl-2 superfamily members of Bcl-2/Bim in CD4 + CD25 − T cells were determined by Western blot analysis. (B) There were no significant differences of Smad2/Smad3 expression in CD4 + CD25 − T cells among various groups; the expressions of P-smad2/P-Smad3 in the CLP group were significantly higher than that in sham group, while P-smad2/P-Smad3 expression in CLP+(PC61 or anti-TGF-β antibody) group was significantly lower than that in the CLP group ( P

    Article Snippet: Rat anti-mouse Bcl-2 and the Bim antibody were purchased from Santa Cruz Biotechnology.

    Techniques: Expressing, Western Blot

    Analysis of B lymphocyte populations in wild-type vs. transgenic mice. Four-color flow-cytometry analysis was performed to determine the phenotype of B lymphocytes. Gating was performed on the B220 + population (R1 and R2, Left ) in the case of CD23/CD21 and IgM/IgD analyses or on the lymphocyte population for B220/CD5 and isotype control analyses. Splenocytes were analyzed from representative 11-month-old wild-type, Bcl-2, TRAF2DN, and TRAF2DN/Bcl-2 double-transgenic mice at a premalignant stage and splenocytes or blood lymphocytes of a representative TRAF2DN/Bcl-2 mouse (12 months old) with acute disease.

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

    Article Title: TNF receptor-associated factor (TRAF) domain and Bcl-2 cooperate to induce small B cell lymphoma/chronic lymphocytic leukemia in transgenic mice

    doi: 10.1073/pnas.0407541101

    Figure Lengend Snippet: Analysis of B lymphocyte populations in wild-type vs. transgenic mice. Four-color flow-cytometry analysis was performed to determine the phenotype of B lymphocytes. Gating was performed on the B220 + population (R1 and R2, Left ) in the case of CD23/CD21 and IgM/IgD analyses or on the lymphocyte population for B220/CD5 and isotype control analyses. Splenocytes were analyzed from representative 11-month-old wild-type, Bcl-2, TRAF2DN, and TRAF2DN/Bcl-2 double-transgenic mice at a premalignant stage and splenocytes or blood lymphocytes of a representative TRAF2DN/Bcl-2 mouse (12 months old) with acute disease.

    Article Snippet: Cell lysates from frozen mouse lymphoid tissues were prepared in modified Laemmli buffer, as described ( ) Tissue lysates were normalized for total protein content (50 μg per lane) and subjected to SDS/PAGE followed by immunoblot analysis using anti-human Bcl-2 , anti-mouse Bcl-2 , anti-TRAF2 (Santa Cruz Biotechnology, C-20), antilung Krüppel-like factor , and anti-Tcl-1 (Santa Cruz Biotechnology, F-14) Abs.

    Techniques: Transgenic Assay, Mouse Assay, Flow Cytometry, Cytometry

    Increased levels of adhesion molecules on the surface of B lymphocytes from TRAF2DN and TRAF2DN/Bcl-2 transgenic mice. ( A ) Surface expression of intercellular adhesion molecule-1 (CD54) and β1-integrin (CD29) was measured on the B220-positive B lymphocytes by flow cytometry. FITC anti-CD29 and PE-anti-CD54 mAbs were used to stain splenic B cells from age-matched wild-type, Bcl-2, TRAF2DN, premalignant TRAF2DN/Bcl-2 double-positive, and TRAF2DN/Bcl-2 double-positive in the acute phase of the disease and blood lymphocytes from this TRAF2DN/Bcl-2 double-positive mouse in the acute phase. ( B ) A similar analysis was performed by using blood from another representative wild-type mouse or lymphocytes from spleen, lungs, blood, or pleural effusion of a TRAF2DN/Bcl-2 mouse that had developed leukemia. ( C ) The percentage of B220-positive lymphocytes with high levels of surface CD54/CD29 expression (upper right quadrant) was compared. B cells from spleen (black dots) or lung (white dots) were analyzed. Each dot represents an individual mouse. Statistical significance was determined by unpaired t test.

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

    Article Title: TNF receptor-associated factor (TRAF) domain and Bcl-2 cooperate to induce small B cell lymphoma/chronic lymphocytic leukemia in transgenic mice

    doi: 10.1073/pnas.0407541101

    Figure Lengend Snippet: Increased levels of adhesion molecules on the surface of B lymphocytes from TRAF2DN and TRAF2DN/Bcl-2 transgenic mice. ( A ) Surface expression of intercellular adhesion molecule-1 (CD54) and β1-integrin (CD29) was measured on the B220-positive B lymphocytes by flow cytometry. FITC anti-CD29 and PE-anti-CD54 mAbs were used to stain splenic B cells from age-matched wild-type, Bcl-2, TRAF2DN, premalignant TRAF2DN/Bcl-2 double-positive, and TRAF2DN/Bcl-2 double-positive in the acute phase of the disease and blood lymphocytes from this TRAF2DN/Bcl-2 double-positive mouse in the acute phase. ( B ) A similar analysis was performed by using blood from another representative wild-type mouse or lymphocytes from spleen, lungs, blood, or pleural effusion of a TRAF2DN/Bcl-2 mouse that had developed leukemia. ( C ) The percentage of B220-positive lymphocytes with high levels of surface CD54/CD29 expression (upper right quadrant) was compared. B cells from spleen (black dots) or lung (white dots) were analyzed. Each dot represents an individual mouse. Statistical significance was determined by unpaired t test.

    Article Snippet: Cell lysates from frozen mouse lymphoid tissues were prepared in modified Laemmli buffer, as described ( ) Tissue lysates were normalized for total protein content (50 μg per lane) and subjected to SDS/PAGE followed by immunoblot analysis using anti-human Bcl-2 , anti-mouse Bcl-2 , anti-TRAF2 (Santa Cruz Biotechnology, C-20), antilung Krüppel-like factor , and anti-Tcl-1 (Santa Cruz Biotechnology, F-14) Abs.

    Techniques: Transgenic Assay, Mouse Assay, Expressing, Flow Cytometry, Cytometry, Staining

    ), and digested with EcoR1 and analyzed by Southern blotting by using a IgH probe. The band of 6 kb corresponds to the germ line (GL) IgH locus. IgH rearrangements corresponding to clonal expansions are indicated with an arrowhead. Wild-type, TRAF2DN, and TRAF2DN/Bcl-2 mice either asymptomatic (premalignant) or with overt lymphoma (disease) were analyzed. The ages in months of the mice (identified by number) analyzed were 17 (102), 15 (137), 14 (158, 157, and 148), 13 (83 and 316), and 12 (302, 327, 87, 89, and 95).

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

    Article Title: TNF receptor-associated factor (TRAF) domain and Bcl-2 cooperate to induce small B cell lymphoma/chronic lymphocytic leukemia in transgenic mice

    doi: 10.1073/pnas.0407541101

    Figure Lengend Snippet: ), and digested with EcoR1 and analyzed by Southern blotting by using a IgH probe. The band of 6 kb corresponds to the germ line (GL) IgH locus. IgH rearrangements corresponding to clonal expansions are indicated with an arrowhead. Wild-type, TRAF2DN, and TRAF2DN/Bcl-2 mice either asymptomatic (premalignant) or with overt lymphoma (disease) were analyzed. The ages in months of the mice (identified by number) analyzed were 17 (102), 15 (137), 14 (158, 157, and 148), 13 (83 and 316), and 12 (302, 327, 87, 89, and 95).

    Article Snippet: Cell lysates from frozen mouse lymphoid tissues were prepared in modified Laemmli buffer, as described ( ) Tissue lysates were normalized for total protein content (50 μg per lane) and subjected to SDS/PAGE followed by immunoblot analysis using anti-human Bcl-2 , anti-mouse Bcl-2 , anti-TRAF2 (Santa Cruz Biotechnology, C-20), antilung Krüppel-like factor , and anti-Tcl-1 (Santa Cruz Biotechnology, F-14) Abs.

    Techniques: Southern Blot, Mouse Assay

    ( A ) Analysis of lymphocyte proliferation. Lymphocytes were isolated from spleens of wild-type (▪), Bcl-2 (♦), TRAF2DN (○), and TRAF2DN/Bcl-2 (Δ) transgenic mice. Cells (5 × 10 4 ) were seeded in triplicate in 96-well plates and pulsed with 1 μCi (1 Ci = 37 GBq) of [ 3 H]-methylthymidine for 12 h at days 0 (immediately after purification), 3, and 6. Proliferation was measured as the mean of [ 3 H]-methylthymidine incorporation (cpm) ± SEM ( n = 3) from three mice of each genotype. ( B ) DNA content analysis; 10 6 lymphocytes from spleen ( a–d ), ascitic fluid ( e ), and lung ( f ) from wild-type ( a ; n = 4), Bcl-2 ( b ; n = 3), TRAF2DN ( c ; n = 4), and symptomatic TRAF2DN/Bcl-2 ( d–f ; n = 4) mice were lysed in hypotonic buffer and stained with PI. Cells with DNA content ≥ of 2n were gated, and the percentage of G 1 cells (DNA content = 2n) and cells in S, G 2 , or M phases ( > 2n) was quantified. Shown are representative DNA content profiles of the various genotypes. The statistical analysis in the text corresponds to mean ± SEM. ( C ) Bcl-2 protects cells from spontaneous apoptosis. Splenocytes isolated from wild-type, Bcl-2, TRAF2DN, and TRAF2DN/Bcl-2 transgenic mice were cultured for 24 h and then incubated with APC-anti-B220 mAb for 1 h, washed, and incubated with annexinV-FITC/PI. B lymphocytes (B220 + cells) were gated, and the average percentage of annexin V-FITC-positive (apoptotic) B lymphocytes and the SD ( n = 3) is shown ( P = 0.01) comparing apoptosis of wild-type vs. Bcl-2 ( * ) or TRAF2DN/Bcl-2 ( ** ). B cells are compared by unpaired t test. ( D ) Effects of chemotherapeutic drugs on apoptosis. Splenocytes isolated from wild-type (▪), Bcl-2 (♦), TRAF2DN (○), and TRAF2DN/Bcl-2 (Δ) transgenic mice were incubated with or without dexamethasone (0.01, 0.1, and 0.5 μM) or F-ara-A (1.25, 2.5, and 5 μM). Cells were harvested after 24 (Dex) or 48 (F-ara-A) h and incubated with APC-anti-B220 mAb for 1 h, then washed and incubated with annexin-V-FITC and PI. B lymphocytes (B220 + cells) were gated, and the percentage of apoptotic cells (annexin V + ) was determined by flow cytometry. Data were corrected for differences in spontaneous apoptosis (mean ± SEM) ( n = 6).

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

    Article Title: TNF receptor-associated factor (TRAF) domain and Bcl-2 cooperate to induce small B cell lymphoma/chronic lymphocytic leukemia in transgenic mice

    doi: 10.1073/pnas.0407541101

    Figure Lengend Snippet: ( A ) Analysis of lymphocyte proliferation. Lymphocytes were isolated from spleens of wild-type (▪), Bcl-2 (♦), TRAF2DN (○), and TRAF2DN/Bcl-2 (Δ) transgenic mice. Cells (5 × 10 4 ) were seeded in triplicate in 96-well plates and pulsed with 1 μCi (1 Ci = 37 GBq) of [ 3 H]-methylthymidine for 12 h at days 0 (immediately after purification), 3, and 6. Proliferation was measured as the mean of [ 3 H]-methylthymidine incorporation (cpm) ± SEM ( n = 3) from three mice of each genotype. ( B ) DNA content analysis; 10 6 lymphocytes from spleen ( a–d ), ascitic fluid ( e ), and lung ( f ) from wild-type ( a ; n = 4), Bcl-2 ( b ; n = 3), TRAF2DN ( c ; n = 4), and symptomatic TRAF2DN/Bcl-2 ( d–f ; n = 4) mice were lysed in hypotonic buffer and stained with PI. Cells with DNA content ≥ of 2n were gated, and the percentage of G 1 cells (DNA content = 2n) and cells in S, G 2 , or M phases ( > 2n) was quantified. Shown are representative DNA content profiles of the various genotypes. The statistical analysis in the text corresponds to mean ± SEM. ( C ) Bcl-2 protects cells from spontaneous apoptosis. Splenocytes isolated from wild-type, Bcl-2, TRAF2DN, and TRAF2DN/Bcl-2 transgenic mice were cultured for 24 h and then incubated with APC-anti-B220 mAb for 1 h, washed, and incubated with annexinV-FITC/PI. B lymphocytes (B220 + cells) were gated, and the average percentage of annexin V-FITC-positive (apoptotic) B lymphocytes and the SD ( n = 3) is shown ( P = 0.01) comparing apoptosis of wild-type vs. Bcl-2 ( * ) or TRAF2DN/Bcl-2 ( ** ). B cells are compared by unpaired t test. ( D ) Effects of chemotherapeutic drugs on apoptosis. Splenocytes isolated from wild-type (▪), Bcl-2 (♦), TRAF2DN (○), and TRAF2DN/Bcl-2 (Δ) transgenic mice were incubated with or without dexamethasone (0.01, 0.1, and 0.5 μM) or F-ara-A (1.25, 2.5, and 5 μM). Cells were harvested after 24 (Dex) or 48 (F-ara-A) h and incubated with APC-anti-B220 mAb for 1 h, then washed and incubated with annexin-V-FITC and PI. B lymphocytes (B220 + cells) were gated, and the percentage of apoptotic cells (annexin V + ) was determined by flow cytometry. Data were corrected for differences in spontaneous apoptosis (mean ± SEM) ( n = 6).

    Article Snippet: Cell lysates from frozen mouse lymphoid tissues were prepared in modified Laemmli buffer, as described ( ) Tissue lysates were normalized for total protein content (50 μg per lane) and subjected to SDS/PAGE followed by immunoblot analysis using anti-human Bcl-2 , anti-mouse Bcl-2 , anti-TRAF2 (Santa Cruz Biotechnology, C-20), antilung Krüppel-like factor , and anti-Tcl-1 (Santa Cruz Biotechnology, F-14) Abs.

    Techniques: Isolation, Transgenic Assay, Mouse Assay, Purification, Staining, Cell Culture, Incubation, Acetylene Reduction Assay, Flow Cytometry, Cytometry

    Histochemical analysis of tissues from the TRAF2DN/Bcl-2 double-transgenic mice. Immunohistochemical analysis of organs and tissues from the TRAF2DN/Bcl-2 double-transgenic mice was performed. Spleen ( A ), prostate ( B ), lung ( C and D ), epididymis ( F and G ), submaxillary gland lymph node ( H ), ascites ( I ), and bone marrow ( J ). Hematoxylin/eosin staining is shown in A–C , E , and G . Staining with anti-B220 mAb is shown in D , F , and I to illustrate that the infiltrating cells are predominately B lymphocytes. Wright–Giemsa staining of ascites cells is shown in H .

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

    Article Title: TNF receptor-associated factor (TRAF) domain and Bcl-2 cooperate to induce small B cell lymphoma/chronic lymphocytic leukemia in transgenic mice

    doi: 10.1073/pnas.0407541101

    Figure Lengend Snippet: Histochemical analysis of tissues from the TRAF2DN/Bcl-2 double-transgenic mice. Immunohistochemical analysis of organs and tissues from the TRAF2DN/Bcl-2 double-transgenic mice was performed. Spleen ( A ), prostate ( B ), lung ( C and D ), epididymis ( F and G ), submaxillary gland lymph node ( H ), ascites ( I ), and bone marrow ( J ). Hematoxylin/eosin staining is shown in A–C , E , and G . Staining with anti-B220 mAb is shown in D , F , and I to illustrate that the infiltrating cells are predominately B lymphocytes. Wright–Giemsa staining of ascites cells is shown in H .

    Article Snippet: Cell lysates from frozen mouse lymphoid tissues were prepared in modified Laemmli buffer, as described ( ) Tissue lysates were normalized for total protein content (50 μg per lane) and subjected to SDS/PAGE followed by immunoblot analysis using anti-human Bcl-2 , anti-mouse Bcl-2 , anti-TRAF2 (Santa Cruz Biotechnology, C-20), antilung Krüppel-like factor , and anti-Tcl-1 (Santa Cruz Biotechnology, F-14) Abs.

    Techniques: Transgenic Assay, Mouse Assay, Immunohistochemistry, Staining

    Characterization of transgenic mice. ( A ) Schematic representation of the Bcl-2 ( Upper ) and TRAF2DN ( Lower ). ( B ) Immunoblot analysis was performed by using lysates from splenocytes isolated from age-matching mice having different genotypes, as indicated. Samples were normalized for protein content and blotted with Abs recognizing mouse TRAF2, human Bcl-2, mouse Bcl-2, lung Krüppel-like factor, and T cell lymphoma-1. +/+, TRAF2DN/Bcl-2 double-transgenic mice. ( C ) Spleen weights are compared from euthanized mice (gray columns) or from mice that died from disease or old age (black columns). The average weight ± SE of spleens from the different transgenic mice was calculated. WT, 138 ± 13.9 mg, n = 10; Bcl-2, 205 ± 6.3 mg, n = 9; TRAF2DN, 496 ± 37.7 mg, n = 10; TRAF2DN/Bcl-2, 647 ± 54 mg, n = 7; TRAF2DN/Bcl-2 dead from disease or old age, 1,247 ± 180 mg, n = 16. * , P

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

    Article Title: TNF receptor-associated factor (TRAF) domain and Bcl-2 cooperate to induce small B cell lymphoma/chronic lymphocytic leukemia in transgenic mice

    doi: 10.1073/pnas.0407541101

    Figure Lengend Snippet: Characterization of transgenic mice. ( A ) Schematic representation of the Bcl-2 ( Upper ) and TRAF2DN ( Lower ). ( B ) Immunoblot analysis was performed by using lysates from splenocytes isolated from age-matching mice having different genotypes, as indicated. Samples were normalized for protein content and blotted with Abs recognizing mouse TRAF2, human Bcl-2, mouse Bcl-2, lung Krüppel-like factor, and T cell lymphoma-1. +/+, TRAF2DN/Bcl-2 double-transgenic mice. ( C ) Spleen weights are compared from euthanized mice (gray columns) or from mice that died from disease or old age (black columns). The average weight ± SE of spleens from the different transgenic mice was calculated. WT, 138 ± 13.9 mg, n = 10; Bcl-2, 205 ± 6.3 mg, n = 9; TRAF2DN, 496 ± 37.7 mg, n = 10; TRAF2DN/Bcl-2, 647 ± 54 mg, n = 7; TRAF2DN/Bcl-2 dead from disease or old age, 1,247 ± 180 mg, n = 16. * , P

    Article Snippet: Cell lysates from frozen mouse lymphoid tissues were prepared in modified Laemmli buffer, as described ( ) Tissue lysates were normalized for total protein content (50 μg per lane) and subjected to SDS/PAGE followed by immunoblot analysis using anti-human Bcl-2 , anti-mouse Bcl-2 , anti-TRAF2 (Santa Cruz Biotechnology, C-20), antilung Krüppel-like factor , and anti-Tcl-1 (Santa Cruz Biotechnology, F-14) Abs.

    Techniques: Transgenic Assay, Mouse Assay, Isolation

    SAR131675 attenuates apoptosis and oxidative stress in the kidneys in  db/db  mice. Glomerulosclerosis and tubulointerstitial fibrosis were determined at 20 weeks in  db/m  and  db/db  mice treated with or without SAR131675.  a  Representative images of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining in the glomeruli and tubules and quantitative data.  b  Representative western blots for B cell leukemia/lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and β-actin in the kidneys and quantitative data.  c  Twenty-four hour urinary 8-hydroxy-deoxyguanosine (8-OH-dG) and isoprostane levels. * P

    Journal: Cell Death & Disease

    Article Title: Inhibition of lymphatic proliferation by the selective VEGFR-3 inhibitor SAR131675 ameliorates diabetic nephropathy in db/db mice

    doi: 10.1038/s41419-019-1436-1

    Figure Lengend Snippet: SAR131675 attenuates apoptosis and oxidative stress in the kidneys in db/db mice. Glomerulosclerosis and tubulointerstitial fibrosis were determined at 20 weeks in db/m and db/db mice treated with or without SAR131675. a Representative images of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining in the glomeruli and tubules and quantitative data. b Representative western blots for B cell leukemia/lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and β-actin in the kidneys and quantitative data. c Twenty-four hour urinary 8-hydroxy-deoxyguanosine (8-OH-dG) and isoprostane levels. * P

    Article Snippet: Western blot analysis Protein extracts were prepared from cultured cells and homogenates of mouse renal cortex or medulla by using mammalian protein extraction reagent (Intron Biotechnology, Gyeonggi-Do, Korea) lysis buffer to which protease inhibitors (Roche, Indianapolis) was added, and Western blots were performed with specific antibodies for VEGF-C (Invitrogen, Carlsbad, CA, USA), VEGFR-1 (Epitomics, Burlingame, CA, USA), VEGFR-2 (Cell Signaling Technology, Danvers, MA, USA) and VEGFR3 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), LYVE-1 (Novus Biologicals, Littleton, CO, USA), podoplanin (ReliaTech GmbH, Wolfenbüttel, Germany), α-SMA (Abcam, Cambridge, UK), fibronectin (ProteinTech, Chicago, IL, USA), inducible nitric oxide synthase (iNOS) (BD Biosciences, San Diego, CA, USA), CD68 (Bio-Rad, Richmond, CA, USA), arginase I (Santa Cruz Biotechnology, Santa Cruz, CA, USA), arginase II (Santa Cruz Biotechnology, Santa Cruz, CA, USA), B cell leukemia/lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and β-actin (Sigma-Aldrich, St. Louis, MO, USA).

    Techniques: Mouse Assay, End Labeling, TUNEL Assay, Staining, Western Blot

    Peptide-induced Ca 2+ oscillations in primary CLL cells. (A) Immunoblot comparing Bcl-2 levels in normal human B lymphocytes and 3 primary CLL samples. (B-D) Representative single-cell Ca 2+ recordings illustrating the Ca 2+ responses observed in CLL cells

    Journal: Blood

    Article Title: Induction of Ca2+-driven apoptosis in chronic lymphocytic leukemia cells by peptide-mediated disruption of Bcl-2-IP3 receptor interaction

    doi: 10.1182/blood-2010-09-307405

    Figure Lengend Snippet: Peptide-induced Ca 2+ oscillations in primary CLL cells. (A) Immunoblot comparing Bcl-2 levels in normal human B lymphocytes and 3 primary CLL samples. (B-D) Representative single-cell Ca 2+ recordings illustrating the Ca 2+ responses observed in CLL cells

    Article Snippet: The following antibodies were used: anti–human Bcl-2 (551052; BD Biosciences), anti–mouse/human Bcl-2 (Santa Cruz Biotechnology), and anti-actin (Sigma).

    Techniques:

    Biochemical and functional characterization of the DD/AA substitution. (A) Typical GST pull-down experiment documenting the binding of 3xFLAG-Bcl-2 in COS-7 cell lysate to GST-Domain3 and GST-Domain3DD/AA using GST as a negative control. GelCode blue–stained

    Journal: Blood

    Article Title: Induction of Ca2+-driven apoptosis in chronic lymphocytic leukemia cells by peptide-mediated disruption of Bcl-2-IP3 receptor interaction

    doi: 10.1182/blood-2010-09-307405

    Figure Lengend Snippet: Biochemical and functional characterization of the DD/AA substitution. (A) Typical GST pull-down experiment documenting the binding of 3xFLAG-Bcl-2 in COS-7 cell lysate to GST-Domain3 and GST-Domain3DD/AA using GST as a negative control. GelCode blue–stained

    Article Snippet: The following antibodies were used: anti–human Bcl-2 (551052; BD Biosciences), anti–mouse/human Bcl-2 (Santa Cruz Biotechnology), and anti-actin (Sigma).

    Techniques: Functional Assay, Binding Assay, Negative Control, Staining

    Bcl-2 dependent induction of Ca 2+ oscillations by TAT-IDP. (A) Diagram of type-1 IP 3 R domains designating the origin of the IDP sequence (blue) and the scrambled control sequence (orange), along with the TAT sequence (gray). (B) Immunoblot documenting

    Journal: Blood

    Article Title: Induction of Ca2+-driven apoptosis in chronic lymphocytic leukemia cells by peptide-mediated disruption of Bcl-2-IP3 receptor interaction

    doi: 10.1182/blood-2010-09-307405

    Figure Lengend Snippet: Bcl-2 dependent induction of Ca 2+ oscillations by TAT-IDP. (A) Diagram of type-1 IP 3 R domains designating the origin of the IDP sequence (blue) and the scrambled control sequence (orange), along with the TAT sequence (gray). (B) Immunoblot documenting

    Article Snippet: The following antibodies were used: anti–human Bcl-2 (551052; BD Biosciences), anti–mouse/human Bcl-2 (Santa Cruz Biotechnology), and anti-actin (Sigma).

    Techniques: Sequencing

    M11L specifically binds to Bak but not to other Bcl-2 family members by use of the TAP method. (A) TAP method for isolating M11L binding partners. The plasmid (M11L-C-TAP) was transiently transfected into 293T cells by the calcium phosphate method (step 1). The lysate with putative TAP M11L binding partner complexes was incubated with rabbit agarose IgG beads in order to bind the protein A of the TAP cassette. The beads were washed to remove unbound proteins and then incubated with TEV protease (step 2) to release bound complexes. The protein complex from cleaved products was allowed to bind via calmodulin binding protein (CBP) resin, and then the purified complex was eluted with calmodulin elution buffer containing EGTA (step 3). The eluants, containing proteins interacting with M11L, were separated by SDS-PAGE, and the proteins were analyzed (step 4). (B) Bak, but not Bad, Bid, Bax/p21, or Bcl-2, was detected as a binding partner of M11L in the M11L-C-TAP eluted products (lane 3), but not in control eluants of C-TAP alone (lane 2), following lysis with CHAPS buffer. Lane 1 confirms endogenous expression levels of the various Bcl-2 family members in 293T cell lysates. (C) Cell lysates of the C-TAP vector alone (lane 1) or the M11L-C-TAP bait (lane 2) were probed with anti-PAP before the TAP procedure to confirm the expression and size of the M11L-TAP fusion protein (34 kDa). (D) Eluants of the C-TAP vector alone (lane 1) or M11L-C-TAP (lane 2) were immunoblotted with anti-M11L, indicating that the M11L bait tagged with CBP (17 kDa) was recovered following TEV cleavage and elution. (E) huBak binds to untagged M11L. Shown are TAP eluants from lysates of cotransfections of pcDNA3-M11L plus N-TAP vector alone (lane 1) or pcDNA3-M11L plus N-TAP-huBak (lane 2) in 293T cells. Lane 3 (M11L and TAP vector) and lane 4 (M11L and N-TAP-huBak) represent corresponding cell lysates, confirming the expression of the appropriate cotransfected plasmids.

    Journal: Journal of Virology

    Article Title: Myxoma Virus M11L Prevents Apoptosis through Constitutive Interaction with Bak

    doi: 10.1128/JVI.78.13.7097-7111.2004

    Figure Lengend Snippet: M11L specifically binds to Bak but not to other Bcl-2 family members by use of the TAP method. (A) TAP method for isolating M11L binding partners. The plasmid (M11L-C-TAP) was transiently transfected into 293T cells by the calcium phosphate method (step 1). The lysate with putative TAP M11L binding partner complexes was incubated with rabbit agarose IgG beads in order to bind the protein A of the TAP cassette. The beads were washed to remove unbound proteins and then incubated with TEV protease (step 2) to release bound complexes. The protein complex from cleaved products was allowed to bind via calmodulin binding protein (CBP) resin, and then the purified complex was eluted with calmodulin elution buffer containing EGTA (step 3). The eluants, containing proteins interacting with M11L, were separated by SDS-PAGE, and the proteins were analyzed (step 4). (B) Bak, but not Bad, Bid, Bax/p21, or Bcl-2, was detected as a binding partner of M11L in the M11L-C-TAP eluted products (lane 3), but not in control eluants of C-TAP alone (lane 2), following lysis with CHAPS buffer. Lane 1 confirms endogenous expression levels of the various Bcl-2 family members in 293T cell lysates. (C) Cell lysates of the C-TAP vector alone (lane 1) or the M11L-C-TAP bait (lane 2) were probed with anti-PAP before the TAP procedure to confirm the expression and size of the M11L-TAP fusion protein (34 kDa). (D) Eluants of the C-TAP vector alone (lane 1) or M11L-C-TAP (lane 2) were immunoblotted with anti-M11L, indicating that the M11L bait tagged with CBP (17 kDa) was recovered following TEV cleavage and elution. (E) huBak binds to untagged M11L. Shown are TAP eluants from lysates of cotransfections of pcDNA3-M11L plus N-TAP vector alone (lane 1) or pcDNA3-M11L plus N-TAP-huBak (lane 2) in 293T cells. Lane 3 (M11L and TAP vector) and lane 4 (M11L and N-TAP-huBak) represent corresponding cell lysates, confirming the expression of the appropriate cotransfected plasmids.

    Article Snippet: Mouse monoclonal antibodies against Bcl-2 (Santa Cruz) and against a peptide epitope derived from the hemagglutinin (HA) protein of human influenza virus (clone 12CA5) (Roche) were also used.

    Techniques: Binding Assay, Plasmid Preparation, Transfection, Incubation, Purification, SDS Page, Lysis, Expressing

    Temporal changes of autophagy and apoptosis after tGCI. a Immunoblots of the apoptosis-related proteins cleaved Caspase-3 and Bcl-2. Total cell lysates from the hippocampus were used, and β-actin was used as an internal control. b Immunoblots of p53, Bax, and cytochrome c in the cytosolic and mitochondrial fractions. GAPDH was used as a loading control for cytosolic proteins, and COX IV was used as the loading control for mitochondrial proteins. c Immunoblots of the autophagy-related proteins LC3B and Beclin-1. Total cell lysates from the hippocampus were used. d – k Densitometric analysis (mean ± SEM, n = 3 animals per group) of the proteins from ( a , b , c ) normalized to the respective loading controls. * P

    Journal: Cell Death & Disease

    Article Title: bFGF plays a neuroprotective role by suppressing excessive autophagy and apoptosis after transient global cerebral ischemia in rats

    doi: 10.1038/s41419-017-0229-7

    Figure Lengend Snippet: Temporal changes of autophagy and apoptosis after tGCI. a Immunoblots of the apoptosis-related proteins cleaved Caspase-3 and Bcl-2. Total cell lysates from the hippocampus were used, and β-actin was used as an internal control. b Immunoblots of p53, Bax, and cytochrome c in the cytosolic and mitochondrial fractions. GAPDH was used as a loading control for cytosolic proteins, and COX IV was used as the loading control for mitochondrial proteins. c Immunoblots of the autophagy-related proteins LC3B and Beclin-1. Total cell lysates from the hippocampus were used. d – k Densitometric analysis (mean ± SEM, n = 3 animals per group) of the proteins from ( a , b , c ) normalized to the respective loading controls. * P

    Article Snippet: The following antibodies were used for immunoblotting: rabbit anti-GAPDH (1:1000, #5174), rabbit anti-p-mTOR (1:1000, #2974), rabbit anti-mTOR (1:1000, #2983), rabbit anti-cleaved Caspase-3 (1:250, #9664), mouse anti-p53 (1:1000, #2524), rabbit anti-Puma (1:200, #7467), rabbit anti-p-4E-BP1 (1:1000, #2855), rabbit anti-4E-BP1 (#9644), rabbit anti-p-p70 S6 (1:1000, #9234), rabbit anti-p70 S6 (1:1000, #2708), rabbit anti-p-ULK1 (1:1000, 14202), rabbit anti-ULK1 (1:1000, #8054), and rabbit anti-COX IV (1:1000, #4850) from Cell Signaling Technology (Danvers, MA, USA); rabbit anti-LC3 (1:2000, ab51520), rabbit anti-p62 (1:1000, ab109012), rabbit anti-cytochrome c (1:2000, ab133504), and rabbit anti-Bax (1:800, ab32503) from Abcam (Cambridge, MA, USA); mouse anti-bFGF (1:1000, sc365106), rabbit anti-Beclin 1 (1:1000, sc-11427), and rabbit anti-Bcl-2 (1:1500, sc-783) from Santa Cruz Biotechnology, Inc. (Santa Cruz CA, USA); and mouse anti-β-actin (1:1000, EM21002), goat anti-rabbit secondary antibody (1:5000, HA1001-100), and goat anti-mouse secondary antibody (1:4000, HA1006) from Hangzhou Huaan Biotechnology (Hangzhou, China).

    Techniques: Western Blot

    Apoptosis was decreased via inhibition of p53 translocation after bFGF treatment. a Immunoblots of cleaved Caspase-3, Bcl-2, and bFGF from total cell lysates of hippocampal tissue harvested 24 h after I/R. β-actin was used as an internal control. b Immunoblots of p53, Puma, and Bax in the cytosolic and mitochondrial fractions of samples harvested 24 h after I/R. β-actin was used as a loading control for cytosolic proteins, and COX IV was used as the loading control for mitochondrial proteins. c – h Densitometric analysis (mean ± SEM, n = 3 animals per group) of the proteins from ( a , b ) were normalized to the respective loading controls. ** P

    Journal: Cell Death & Disease

    Article Title: bFGF plays a neuroprotective role by suppressing excessive autophagy and apoptosis after transient global cerebral ischemia in rats

    doi: 10.1038/s41419-017-0229-7

    Figure Lengend Snippet: Apoptosis was decreased via inhibition of p53 translocation after bFGF treatment. a Immunoblots of cleaved Caspase-3, Bcl-2, and bFGF from total cell lysates of hippocampal tissue harvested 24 h after I/R. β-actin was used as an internal control. b Immunoblots of p53, Puma, and Bax in the cytosolic and mitochondrial fractions of samples harvested 24 h after I/R. β-actin was used as a loading control for cytosolic proteins, and COX IV was used as the loading control for mitochondrial proteins. c – h Densitometric analysis (mean ± SEM, n = 3 animals per group) of the proteins from ( a , b ) were normalized to the respective loading controls. ** P

    Article Snippet: The following antibodies were used for immunoblotting: rabbit anti-GAPDH (1:1000, #5174), rabbit anti-p-mTOR (1:1000, #2974), rabbit anti-mTOR (1:1000, #2983), rabbit anti-cleaved Caspase-3 (1:250, #9664), mouse anti-p53 (1:1000, #2524), rabbit anti-Puma (1:200, #7467), rabbit anti-p-4E-BP1 (1:1000, #2855), rabbit anti-4E-BP1 (#9644), rabbit anti-p-p70 S6 (1:1000, #9234), rabbit anti-p70 S6 (1:1000, #2708), rabbit anti-p-ULK1 (1:1000, 14202), rabbit anti-ULK1 (1:1000, #8054), and rabbit anti-COX IV (1:1000, #4850) from Cell Signaling Technology (Danvers, MA, USA); rabbit anti-LC3 (1:2000, ab51520), rabbit anti-p62 (1:1000, ab109012), rabbit anti-cytochrome c (1:2000, ab133504), and rabbit anti-Bax (1:800, ab32503) from Abcam (Cambridge, MA, USA); mouse anti-bFGF (1:1000, sc365106), rabbit anti-Beclin 1 (1:1000, sc-11427), and rabbit anti-Bcl-2 (1:1500, sc-783) from Santa Cruz Biotechnology, Inc. (Santa Cruz CA, USA); and mouse anti-β-actin (1:1000, EM21002), goat anti-rabbit secondary antibody (1:5000, HA1001-100), and goat anti-mouse secondary antibody (1:4000, HA1006) from Hangzhou Huaan Biotechnology (Hangzhou, China).

    Techniques: Inhibition, Translocation Assay, Western Blot

    Immunohistochemical staining and western blotting of proliferation, apoptosis, and migration/invasion by CoQ 0 in B16F10 xenografted tumors A. In situ apoptosis detection using TUNEL staining in tumor sections from control animals and experimental analogues treated with CoQ 0 (2 mg/kg). Arrow indicates example apoptotic-positive cells (400 × magnifications). The number of apoptotic-positive cells in microscopic fields from 3 samples was averaged. B. Xenografted tumor sections were subjected to immunohistochemical analysis for β-catenin, cyclin D1, survivin, and MMP-9. Cells positive for β-catenin, cyclin D1, survivin, and MMP-9 were counted from 3 fields (200× magnification) for each tumor sample. The number of positive cells (arrows indicate proliferating cells) in microscopic fields from 5∼7 samples was averaged. Results are the mean (±SE) number of cells/microscope field (as percentage) for 5∼7 animals per group. C. Western blotting results showing the effects of CoQ 0 on the total protein contents of β-catenin, c-myc, cyclin D1, survivin, p53, proPARP, Bcl-2, and Bax in the xenografted tumors from 3 samples. Relative changes in protein bands were measured by densitometric analysis with the control being 100% as shown just below the gel data. Significant at * p

    Journal: Oncotarget

    Article Title: In vitro and in vivo anti-tumor activity of CoQ0 against melanoma cells: inhibition of metastasis and induction of cell-cycle arrest and apoptosis through modulation of Wnt/β-catenin signaling pathways

    doi: 10.18632/oncotarget.7983

    Figure Lengend Snippet: Immunohistochemical staining and western blotting of proliferation, apoptosis, and migration/invasion by CoQ 0 in B16F10 xenografted tumors A. In situ apoptosis detection using TUNEL staining in tumor sections from control animals and experimental analogues treated with CoQ 0 (2 mg/kg). Arrow indicates example apoptotic-positive cells (400 × magnifications). The number of apoptotic-positive cells in microscopic fields from 3 samples was averaged. B. Xenografted tumor sections were subjected to immunohistochemical analysis for β-catenin, cyclin D1, survivin, and MMP-9. Cells positive for β-catenin, cyclin D1, survivin, and MMP-9 were counted from 3 fields (200× magnification) for each tumor sample. The number of positive cells (arrows indicate proliferating cells) in microscopic fields from 5∼7 samples was averaged. Results are the mean (±SE) number of cells/microscope field (as percentage) for 5∼7 animals per group. C. Western blotting results showing the effects of CoQ 0 on the total protein contents of β-catenin, c-myc, cyclin D1, survivin, p53, proPARP, Bcl-2, and Bax in the xenografted tumors from 3 samples. Relative changes in protein bands were measured by densitometric analysis with the control being 100% as shown just below the gel data. Significant at * p

    Article Snippet: Anti-rabbit MMP-2, anti-goat MMP-9, anti-mouse Bax, anti-mouse β-actin, anti-rabbit c-myc, anti-rabbit survivin, anti-rabbit Bcl-2, anti-mouse β-catenin, anti-rabbit p53, anti-rabbit caspase-3, antibodies were purchased from Santa Cruz Biotechnology, Inc. (Heidelberg, Germany).

    Techniques: Immunohistochemistry, Staining, Western Blot, Migration, In Situ, TUNEL Assay, Microscopy

    CoQ 0 induced apoptosis and G1 cell-cycle arrest in melanoma B16F10 cells Cells exposed to CoQ 0 (0-20 μM) for 24 h. A. TUNEL assay was performed to determine CoQ 0 -induced apoptosis by directly measuring DNA fragmentation. A histogram indicates the percentage of apoptotic-positive cells induced by CoQ 0 . B. Cells were stained with Annexin V and PI, and analyzed for apoptosis using flow cytometry. Representative flow cytometry patterns are shown. C-E. Western blot analysis was performed to measure the expression levels of apoptotic- and cell cycle-related proteins. The effects of CoQ 0 on the protein levels of Bcl-2, Bax (C), procaspase-3/-9, pro-PARP (D), Cyclin E, CDK4, and p53 (E) in B16F10 cells were monitored with specific antibodies. The results are presented as the mean ± S.D of three independent assays. Significant at * p

    Journal: Oncotarget

    Article Title: In vitro and in vivo anti-tumor activity of CoQ0 against melanoma cells: inhibition of metastasis and induction of cell-cycle arrest and apoptosis through modulation of Wnt/β-catenin signaling pathways

    doi: 10.18632/oncotarget.7983

    Figure Lengend Snippet: CoQ 0 induced apoptosis and G1 cell-cycle arrest in melanoma B16F10 cells Cells exposed to CoQ 0 (0-20 μM) for 24 h. A. TUNEL assay was performed to determine CoQ 0 -induced apoptosis by directly measuring DNA fragmentation. A histogram indicates the percentage of apoptotic-positive cells induced by CoQ 0 . B. Cells were stained with Annexin V and PI, and analyzed for apoptosis using flow cytometry. Representative flow cytometry patterns are shown. C-E. Western blot analysis was performed to measure the expression levels of apoptotic- and cell cycle-related proteins. The effects of CoQ 0 on the protein levels of Bcl-2, Bax (C), procaspase-3/-9, pro-PARP (D), Cyclin E, CDK4, and p53 (E) in B16F10 cells were monitored with specific antibodies. The results are presented as the mean ± S.D of three independent assays. Significant at * p

    Article Snippet: Anti-rabbit MMP-2, anti-goat MMP-9, anti-mouse Bax, anti-mouse β-actin, anti-rabbit c-myc, anti-rabbit survivin, anti-rabbit Bcl-2, anti-mouse β-catenin, anti-rabbit p53, anti-rabbit caspase-3, antibodies were purchased from Santa Cruz Biotechnology, Inc. (Heidelberg, Germany).

    Techniques: TUNEL Assay, Staining, Flow Cytometry, Cytometry, Western Blot, Expressing

    Effect of EGCG on protein expression of apoptotic markers Bax and Bcl-2 in kidney of control and fluoride-treated rats. Protein expression was determined by Western blot analyses. Values are expressed as mean ± SD for measurements from six rats in each group. Statistical significance was determined by one way ANOVA followed by post hoc t -test. Bars with different superscript letters (a–c) differ significantly at P

    Journal: Toxicology Reports

    Article Title: Epigallocatechin gallate supplementation protects against renal injury induced by fluoride intoxication in rats: Role of Nrf2/HO-1 signaling

    doi: 10.1016/j.toxrep.2014.01.002

    Figure Lengend Snippet: Effect of EGCG on protein expression of apoptotic markers Bax and Bcl-2 in kidney of control and fluoride-treated rats. Protein expression was determined by Western blot analyses. Values are expressed as mean ± SD for measurements from six rats in each group. Statistical significance was determined by one way ANOVA followed by post hoc t -test. Bars with different superscript letters (a–c) differ significantly at P

    Article Snippet: After blockage of nonspecific binding sites with 5% nonfat milk in PBS-T (PBS and 1% Tween 20) for 1 h at room temperature, the membrane was incubated overnight at 4 °C with diluted goat anti-Bcl-2, Bax, cytochrome c, cleaved caspases-3 and -9, or β-actin monoclonal antibodies (Santa Cruz Biotechnology, Inc.).

    Techniques: Expressing, Western Blot

    RCAS-Bcl-2 decreases the apoptotic index in anaplastic oligodendroglioma

    Journal: International Journal of Cancer. Journal International du Cancer

    Article Title: Bcl-2 promotes malignant progression in a PDGF-B-dependent murine model of oligodendroglioma

    doi: 10.1002/ijc.25869

    Figure Lengend Snippet: RCAS-Bcl-2 decreases the apoptotic index in anaplastic oligodendroglioma

    Article Snippet: Bcl-2 expression induced by RCAS-Bcl-2 was verified in tumor-bearing sections by immunohistochemical staining for a human Bcl-2 specific antibody Mab100 but does not detect endogenous mouse Bcl-2 ( ).

    Techniques:

    RCAS-Bcl-2 decreases the latency of symptomatic tumor formation

    Journal: International Journal of Cancer. Journal International du Cancer

    Article Title: Bcl-2 promotes malignant progression in a PDGF-B-dependent murine model of oligodendroglioma

    doi: 10.1002/ijc.25869

    Figure Lengend Snippet: RCAS-Bcl-2 decreases the latency of symptomatic tumor formation

    Article Snippet: Bcl-2 expression induced by RCAS-Bcl-2 was verified in tumor-bearing sections by immunohistochemical staining for a human Bcl-2 specific antibody Mab100 but does not detect endogenous mouse Bcl-2 ( ).

    Techniques:

    RCAS-Bcl-2 increases cellular proliferation in tumors

    Journal: International Journal of Cancer. Journal International du Cancer

    Article Title: Bcl-2 promotes malignant progression in a PDGF-B-dependent murine model of oligodendroglioma

    doi: 10.1002/ijc.25869

    Figure Lengend Snippet: RCAS-Bcl-2 increases cellular proliferation in tumors

    Article Snippet: Bcl-2 expression induced by RCAS-Bcl-2 was verified in tumor-bearing sections by immunohistochemical staining for a human Bcl-2 specific antibody Mab100 but does not detect endogenous mouse Bcl-2 ( ).

    Techniques:

    Endogenous and Ectopic Bcl-2 expression in low-grade and anaplastic oligodendrogliomas (scale bar = 50 μm)

    Journal: International Journal of Cancer. Journal International du Cancer

    Article Title: Bcl-2 promotes malignant progression in a PDGF-B-dependent murine model of oligodendroglioma

    doi: 10.1002/ijc.25869

    Figure Lengend Snippet: Endogenous and Ectopic Bcl-2 expression in low-grade and anaplastic oligodendrogliomas (scale bar = 50 μm)

    Article Snippet: Bcl-2 expression induced by RCAS-Bcl-2 was verified in tumor-bearing sections by immunohistochemical staining for a human Bcl-2 specific antibody Mab100 but does not detect endogenous mouse Bcl-2 ( ).

    Techniques: Expressing

    RCAS-Bcl-2 promotes malignancy in the RCAS-PDGFB-dependent model of oligodendroglioma

    Journal: International Journal of Cancer. Journal International du Cancer

    Article Title: Bcl-2 promotes malignant progression in a PDGF-B-dependent murine model of oligodendroglioma

    doi: 10.1002/ijc.25869

    Figure Lengend Snippet: RCAS-Bcl-2 promotes malignancy in the RCAS-PDGFB-dependent model of oligodendroglioma

    Article Snippet: Bcl-2 expression induced by RCAS-Bcl-2 was verified in tumor-bearing sections by immunohistochemical staining for a human Bcl-2 specific antibody Mab100 but does not detect endogenous mouse Bcl-2 ( ).

    Techniques:

    Proposed model of targeting therapy and signaling pathway involved in CGOML-induced apoptosis of MCF-7 cells: Nude mice bearing breast cancer (MCF-7) received intravenous injections of GML (35 μg of Gemcitabine/g) at day 1, 5 and then these mice also received intravenous injections of OML (5 μg of oxaliplatin/g) at day 3, 7; magnetic field (5000 GS) was applied to the tumor surface for 30 min after every injection The magnetic properties of GML or OML particles guide the gemcitabine or oxaliplatin to the tumor area of nude mice under an external magnetic field (Nd 2 Fe 12 B magnet tablets). GML or OML has excellent half-life periods and can be gradually biodegraded in MCF-7 cells, lead to gemcitabine of GML (or oxaliplatin of OML) slowly released in target cells. In MCF-7 cells, OML inhibited the replication and transcription of DNA by forming inter-strand and intra-strand platinum-DNA adducts (Pt-GG and Pt-AG), while GML prevented the synthesis and repair of DNA by inhibiting activation of ribonucleotide reductase, and stopped the the synthesis of DNA and broken strands of DNA by replacing the cytidine of DNA strands with dFdCTP. It leads to the phenomenon of DNA ladder, increase of BAX and decrease of Bcl-2 and Survivin, eventual activation of caspase-9 and caspase-3 cause cell apoptosis in MCF-7 cells, resulting in MCF-7 cell death.

    Journal: Oncotarget

    Article Title: Combination of gemcitabine-containing magnetoliposome and oxaliplatin-containing magnetoliposome in breast cancer treatment: A possible mechanism with potential for clinical application

    doi: 10.18632/oncotarget.9671

    Figure Lengend Snippet: Proposed model of targeting therapy and signaling pathway involved in CGOML-induced apoptosis of MCF-7 cells: Nude mice bearing breast cancer (MCF-7) received intravenous injections of GML (35 μg of Gemcitabine/g) at day 1, 5 and then these mice also received intravenous injections of OML (5 μg of oxaliplatin/g) at day 3, 7; magnetic field (5000 GS) was applied to the tumor surface for 30 min after every injection The magnetic properties of GML or OML particles guide the gemcitabine or oxaliplatin to the tumor area of nude mice under an external magnetic field (Nd 2 Fe 12 B magnet tablets). GML or OML has excellent half-life periods and can be gradually biodegraded in MCF-7 cells, lead to gemcitabine of GML (or oxaliplatin of OML) slowly released in target cells. In MCF-7 cells, OML inhibited the replication and transcription of DNA by forming inter-strand and intra-strand platinum-DNA adducts (Pt-GG and Pt-AG), while GML prevented the synthesis and repair of DNA by inhibiting activation of ribonucleotide reductase, and stopped the the synthesis of DNA and broken strands of DNA by replacing the cytidine of DNA strands with dFdCTP. It leads to the phenomenon of DNA ladder, increase of BAX and decrease of Bcl-2 and Survivin, eventual activation of caspase-9 and caspase-3 cause cell apoptosis in MCF-7 cells, resulting in MCF-7 cell death.

    Article Snippet: Antibodies, namely, rabbit anti-human Bcl-2, BAX, survivin, GAPDH, and goat anti-rabbit IgG-HRP, were purchased from Santa Cruz Biotechnology (Santa Cruz, USA).

    Techniques: Mouse Assay, Injection, Activation Assay

    CGOML activate cell apoptosis in nude mice bearing breast cancer (MCF-7) Notes: A. RT-qPCR results show the mRNA levels of BAX, Bcl-2 and Survivin in tumors collected from mice of 5 different groups (Control, CGO, CGOL, ML, and CGOML groups). The left panel exhibits the relative mRNA levels of BAX in the 5 groups, the middle panel exhibits the relative mRNA levels of Bcl-2 in the 5 groups, whereas the right panel exhibits the relative mRNA levels of Survivin in the 5 groups. B. The mRNA level of BAX/Bcl-2 ratio was determined from results of (A) in each group. C. Western blot analysis shows BAX-, Bcl-2- and Survivin protein levels in tumor tissues collected from nude mice in different groups (Control, CGO, CGOL, ML, and CGOML). D. The protein levels of BAX/Bcl-2 ratio was calculated from results of (C). * p

    Journal: Oncotarget

    Article Title: Combination of gemcitabine-containing magnetoliposome and oxaliplatin-containing magnetoliposome in breast cancer treatment: A possible mechanism with potential for clinical application

    doi: 10.18632/oncotarget.9671

    Figure Lengend Snippet: CGOML activate cell apoptosis in nude mice bearing breast cancer (MCF-7) Notes: A. RT-qPCR results show the mRNA levels of BAX, Bcl-2 and Survivin in tumors collected from mice of 5 different groups (Control, CGO, CGOL, ML, and CGOML groups). The left panel exhibits the relative mRNA levels of BAX in the 5 groups, the middle panel exhibits the relative mRNA levels of Bcl-2 in the 5 groups, whereas the right panel exhibits the relative mRNA levels of Survivin in the 5 groups. B. The mRNA level of BAX/Bcl-2 ratio was determined from results of (A) in each group. C. Western blot analysis shows BAX-, Bcl-2- and Survivin protein levels in tumor tissues collected from nude mice in different groups (Control, CGO, CGOL, ML, and CGOML). D. The protein levels of BAX/Bcl-2 ratio was calculated from results of (C). * p

    Article Snippet: Antibodies, namely, rabbit anti-human Bcl-2, BAX, survivin, GAPDH, and goat anti-rabbit IgG-HRP, were purchased from Santa Cruz Biotechnology (Santa Cruz, USA).

    Techniques: Mouse Assay, Quantitative RT-PCR, Western Blot

    Magnetoliposomes (MLs) containing recombinant human IFNα 2 b (MIL) activate cell apoptosis in liver tumors. Notes: ( A ) Quantitative real-time reverse-transcription polymerase chain-reaction results show the transcription levels of Bax and Bcl-2 in tumor samples obtained from mice of five different groups (saline [NS], IFNα 2 b [IFN], IFNα 2 b liposome [IL], ML, and MIL groups). The right panel shows the relative messenger ribonucleic acid (mRNA) levels of Bax in the five groups, whereas the left panel shows the relative mRNA levels of Bcl-2 in the five groups. ( B ) Ratio of Bax/Bcl-2-transcription levels calculated from results of ( A ) in each group. ( C ) Western blot analysis shows Bax- and Bcl-2-expression levels in tumor tissues obtained from nude mice in different groups (NS, IFN, IL, ML, and MIL). ( D ) Ratio of Bax/Bcl-2 protein levels in tumor tissues of nude mice among the different groups, as determined from results of ( C ). *Significant difference when compared with the NS group ( P

    Journal: International Journal of Nanomedicine

    Article Title: Preclinical evaluation of recombinant human IFNα2b-containing magnetoliposomes for treating hepatocellular carcinoma

    doi: 10.2147/IJN.S67228

    Figure Lengend Snippet: Magnetoliposomes (MLs) containing recombinant human IFNα 2 b (MIL) activate cell apoptosis in liver tumors. Notes: ( A ) Quantitative real-time reverse-transcription polymerase chain-reaction results show the transcription levels of Bax and Bcl-2 in tumor samples obtained from mice of five different groups (saline [NS], IFNα 2 b [IFN], IFNα 2 b liposome [IL], ML, and MIL groups). The right panel shows the relative messenger ribonucleic acid (mRNA) levels of Bax in the five groups, whereas the left panel shows the relative mRNA levels of Bcl-2 in the five groups. ( B ) Ratio of Bax/Bcl-2-transcription levels calculated from results of ( A ) in each group. ( C ) Western blot analysis shows Bax- and Bcl-2-expression levels in tumor tissues obtained from nude mice in different groups (NS, IFN, IL, ML, and MIL). ( D ) Ratio of Bax/Bcl-2 protein levels in tumor tissues of nude mice among the different groups, as determined from results of ( C ). *Significant difference when compared with the NS group ( P

    Article Snippet: Polyclonal antibodies of rabbit antihuman Bcl-2, Bax, and β-actin were purchased from Santa Cruz Biotech (USA).

    Techniques: Recombinant, Reverse Transcription Polymerase Chain Reaction, Mouse Assay, Western Blot, Expressing

    Western blot analysis of Bax, Bcl-2 and Bcl-XL protein expression levels in resveratrol-treated HeLa cells. (A) Cells were exposed to 0, 5, 10, 20 and 40 µmol/l of resveratrol for 48 h. (B) Cells were treated with 20 µmol/l resveratrol for 0, 12, 24, 36 and 48 h. All cells were lysed, equal quantities of protein were separated using 12% SDS-PAGE and protein expression was detected by western blotting. Bcl-2, B-cell lymphoma 2; Bax, B-cell lymphoma 2-associated X protein; Bcl-2 XL, B-cell lymphoma 2-extra large.

    Journal: Oncology Letters

    Article Title: Resveratrol suppresses human cervical carcinoma cell proliferation and elevates apoptosis via the mitochondrial and p53 signaling pathways

    doi: 10.3892/ol.2018.8571

    Figure Lengend Snippet: Western blot analysis of Bax, Bcl-2 and Bcl-XL protein expression levels in resveratrol-treated HeLa cells. (A) Cells were exposed to 0, 5, 10, 20 and 40 µmol/l of resveratrol for 48 h. (B) Cells were treated with 20 µmol/l resveratrol for 0, 12, 24, 36 and 48 h. All cells were lysed, equal quantities of protein were separated using 12% SDS-PAGE and protein expression was detected by western blotting. Bcl-2, B-cell lymphoma 2; Bax, B-cell lymphoma 2-associated X protein; Bcl-2 XL, B-cell lymphoma 2-extra large.

    Article Snippet: P53 antibody (cat. no. 38007) was purchased from Signalway Antibody (College Park, MA, USA) and B-cell lymphoma 2 (Bcl-2) (cat. no. sc-7382), Bcl-2-associated X protein (Bax) (cat. no. sc-7480), Bcl-extra large (XL; cat. no. sc-8392), caspase-3 (cat. no. sc-7272), β-actin (cat. no. sc-8432) and were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).

    Techniques: Western Blot, Expressing, SDS Page