anti-cleaved caspase-3 Search Results


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  • 98
    Millipore anti cleaved caspase 3
    SeP KO mice reduces apoptotic cells after myocardial I/R. LV tissue sections were subjected to TUNEL and 4′,6-diamidino-2-phenylindole (DAPI) staining. ( A ) Representative examples of TUNEL-positive myocytes in the ischemic area. Arrows indicate TUNEL-positive myocytes. ( B ) The number of TUNEL-positive myocytes was expressed as a percentage of total nuclei detected using DAPI staining. Heart homogenates were prepared from SeP KO and WT mice subjected to 30 min of ischemia and 24 h of reperfusion. Immunoblot analyses were performed using ( C ) anti-cleaved <t>caspase-3</t> and GAPDH antibody. ( D ) Results of quantitative analysis of cleaved caspase-3 are shown. Data represent means from at least five mice each. * p
    Anti Cleaved Caspase 3, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 203 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Biocare Medical anti cleaved caspase 3 casp3
    Effect of ALLO on the apoptotic index (cleaved <t>CASP3</t> positive immunostained cells over the total number of cells) in follicles ( a ) and corpora lutea ( b ). Control groups follicles (1A upper panel), ALLO treated follicles (1A lower panel), control groups CL (1B upper panel), ALLO treated groups CL (1B lower panel); E, D1, D2, PE from left to right. Representative photomicrographs of follicles and corpora lutea immunostained for cleaved CASP3 at each stage of the estrous cycle. Values are expressed as mean ± S.E.M. Two-way ANOVA followed by Bonferroni’s post hoc, ( n =6), * p
    Anti Cleaved Caspase 3 Casp3, supplied by Biocare Medical, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson anti cleaved caspase 3
    BTLA signaling motifs show no effect on tumor killing, while Grb2 motif augments IL-2 production and T cell proliferation ( a ) Schematic diagram depicts the structure of BTLA WT (left), BTLAΔGrb2 (middle), and BTLAΔITSM (right). Signaling motifs with modified Tyrosine to Phenylalanine are indicated by a dotted pattern over a black background. A table summarizing the phenotype observed with the expression of BTLA or of the different BTLA constructs is presented on the right panel. ( b ) B16 OVA (mouse melanoma tumor positive for OVA) or B16F10 (mouse melanoma tumor negative for OVA) were stained with eFluor670® and co-cultured with OT-1 BTLA KO T cells overexpressing WT BTLA or BTLA mutants at the following T cell-to-tumor cell ratios (1:10, 1:3, and 1:1). Tumor cell death is depicted by the percentage of <t>caspase-3</t> positive cells. N=3 independent experiments ( c ) OT-1 BTLA KO T cells overexpressing WT BTLA or its variants were re-stimulated with dendritic cells pulsed with OVA peptide. TNF-α and IFN-γ production by virally transduced OT-1 BTLA KO T cells was evaluated by intracellular staining. Bar graph depicts the percentage of positive cells (left panel) and mean fluorescence intensity (MFI) (right panel). Each bar represents three independent experiments. (Two-way ANOVA; * P
    Anti Cleaved Caspase 3, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 148 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology anti cleaved caspase 3
    Expression of p53 is enhanced by miR-128 mimics via inhibition of MDM4 expression. (A) Western blot analysis of MDM4, p53 and cleaved <t>caspase-3</t> expression following transfection with miR-128 mimics. (B) Quantification of MDM4 expression. (C) Quantification of p53 expression. (D) Quantification of cleaved caspase-3 expression. **P
    Anti Cleaved Caspase 3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 487 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Merck KGaA anti cleaved caspase 3
    P-MVs-secreted miR-191 affected tubular epithelial cell apoptosis via CBS. P-MVs isolated from patients with renal allograft rejection were divided into seven groups: control, P-MVs of healthy, P-MVs of patients, P-MVs of patients+NC, P-MVs of patients+miR-191 Inhibitor, P-MVs of patients+miR-191 inhibitor+si-control, P-MVs of patients+miR-191 Inhibitor+si-CBS. A. HK-2 cell apoptosis was observed by flow cytometry. B. Apoptosis related proteins of Bax, Bcl-2, <t>Caspase</t> 3 were measured. *P
    Anti Cleaved Caspase 3, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 93/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    R&D Systems anti cleaved caspase 3
    Cells derived from Ela-myc pancreatic tumors are sensitive to sunitinib in vitro A. Analysis by Western blot of the expresssion of the sunitinib-targeted receptors VEGFR2, PDGFR-α and PDGFR–β in two different cell lines derived from Ela-myc pancreatic tumors. B. Bright field images showing Emyc-1 cell sensitivity to different doses of sunitinib (1, 2 or 4 μM) in vitro , as compared to cells treated with vehicle (DMSO). Scale bars, 100 μm. C. MTT experiments were performed to quantify cell viability upon sunitinib treatment (1, 2 or 4 μM) in the Emyc-1 cell line, observing a dose-dependent effect. D. Left, immunofluorescence of cleaved caspase 3, to detect cell apoptosis upon sunitinib treatment (1, 2 or 4 μM). Scale bars, 20 μm. Right, Western blot analysis of the levels of cleaved <t>caspase</t> 3 in Emyc-1 cell line treated with sunitinib. Tubulin levels are shown as the loading control. E. Left, Immunofluorescence of P-Histone H3, to show cell growth arrest upon sunitinib treatment (1, 2 or 4 μM) in Emyc-1 cells. Scale bars, 50 μm. Right, Bar plots showing quantification of P-Histone H3 immunofluorescence experiments on the right. * p
    Anti Cleaved Caspase 3, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 66 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Signalway Antibody anti cleaved caspase 3
    CRTH 2 deletion reduces doxorubicin‐induced cardiomyocyte apoptosis and cardiac injury in mice Western blot analysis of ER stress markers p‐IRE1 and cleaved ATF6 in mouse cardiomyocytes upon DOX (1 μmol/l) treatment. CRTH2 mRNA expression in mouse cardiomyocytes under DOX treatment. Data represent mean ± SEM. ** P = 0.00011, 12 h vs. 0 h; ** P = 0.000267, 24 h vs. 0 h (one‐way ANOVA); n = 4. Cardiac function in mice assessed by M‐mode echocardiographic analysis on day 7 after DOX treatment (20 mg/kg, i.p.). EF, ejection fraction (C); FS, fractional shortening (D); LVPWs, left ventricle posterior wall thickness at end‐systole (E). Data represent mean ± SEM. EF, * P = 0.00836, vs. WT; FS, * P = 0.00383, vs. WT; LVPWs, * P = 0.0198, vs. WT (unpaired two‐tailed t ‐test); WT, n = 15, CRTH2 −/− , n = 13. Kaplan–Meier survival curves for mice subjected to DOX treatment. * P = 0.0305, vs. WT (log‐rank test); n = 15. Representative TUNEL‐stained images of heart tissue in mice on day 7 after DOX treatment. Green, TUNEL‐positive nuclei; blue, DAPI; red, α‐actinin; scale bar, 50 μm. Quantification of TUNEL‐positive cardiomyocytes in (G). Data represent mean ± SEM. * P = 0.00278, vs. WT (unpaired two‐tailed t ‐test); n = 6. Western blot analysis of m‐calpain, caspase‐12, and <t>caspase‐3</t> in heart tissue from DOX‐treated mice. Schematic diagram of CRTH2 promoting cardiomyocyte apoptosis under ER stress through the G αq /calpain/caspase‐12 signaling pathway. Source data are available online for this figure.
    Anti Cleaved Caspase 3, supplied by Signalway Antibody, used in various techniques. Bioz Stars score: 92/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Trevigen anti cleaved caspase 3
    CRTH 2 deletion reduces doxorubicin‐induced cardiomyocyte apoptosis and cardiac injury in mice Western blot analysis of ER stress markers p‐IRE1 and cleaved ATF6 in mouse cardiomyocytes upon DOX (1 μmol/l) treatment. CRTH2 mRNA expression in mouse cardiomyocytes under DOX treatment. Data represent mean ± SEM. ** P = 0.00011, 12 h vs. 0 h; ** P = 0.000267, 24 h vs. 0 h (one‐way ANOVA); n = 4. Cardiac function in mice assessed by M‐mode echocardiographic analysis on day 7 after DOX treatment (20 mg/kg, i.p.). EF, ejection fraction (C); FS, fractional shortening (D); LVPWs, left ventricle posterior wall thickness at end‐systole (E). Data represent mean ± SEM. EF, * P = 0.00836, vs. WT; FS, * P = 0.00383, vs. WT; LVPWs, * P = 0.0198, vs. WT (unpaired two‐tailed t ‐test); WT, n = 15, CRTH2 −/− , n = 13. Kaplan–Meier survival curves for mice subjected to DOX treatment. * P = 0.0305, vs. WT (log‐rank test); n = 15. Representative TUNEL‐stained images of heart tissue in mice on day 7 after DOX treatment. Green, TUNEL‐positive nuclei; blue, DAPI; red, α‐actinin; scale bar, 50 μm. Quantification of TUNEL‐positive cardiomyocytes in (G). Data represent mean ± SEM. * P = 0.00278, vs. WT (unpaired two‐tailed t ‐test); n = 6. Western blot analysis of m‐calpain, caspase‐12, and <t>caspase‐3</t> in heart tissue from DOX‐treated mice. Schematic diagram of CRTH2 promoting cardiomyocyte apoptosis under ER stress through the G αq /calpain/caspase‐12 signaling pathway. Source data are available online for this figure.
    Anti Cleaved Caspase 3, supplied by Trevigen, used in various techniques. Bioz Stars score: 93/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Proteintech anti cleaved caspase 3
    Effects of PCNP on the apoptosis of human neuroblastoma cells. a The apoptotic levels of SH-SY5Y and SK-N-SH cells were measured by TUNEL staining; original magnification 100×. b The percentages of TUNEL-positive cells were calculated by the formula shown above. c Western blotting analysis for the expression of cleaved <t>caspase-3,</t> − 8, and − 9 and cleaved PARP in SH-SY5Y and SK-N-SH cells. GAPDH was used as the loading control. ( d, e ) The densitometry analysis of each factor was performed in SH-SY5Y and SK-N-SH cells, normalized to the corresponding GAPDH level. Data are presented as mean ± SEM of three independent experiments; * P
    Anti Cleaved Caspase 3, supplied by Proteintech, used in various techniques. Bioz Stars score: 92/100, based on 90 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti cleaved caspase 3 casp3
    Oligodendrocyte precursor cell susceptibility to apoptotic induction following inflammatory stress. Analysis of <t>Casp3</t> expression in O4+ cells following an inflammatory stress. The fold change of apoptotic (Casp3+) and viable (Casp3-) O4+ cells was similar in the WT and Hp70.1 KO CNS cell cultures after 24 h of stimulation with LPS plus IFN-γ. The error bars correspond to the SEM.
    Anti Cleaved Caspase 3 Casp3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Servicebio Inc anti cleaved caspase 3
    Upregulating H3K27me3 levels sensitizes OS to cisplatin in vivo.  a  Macroscopic image of tumor size after mice were sacrificed. Tumor volume ( b ) and weight ( c ) were measured at the indicated times.  d  H  E, TUNEL, cleaved Caspase 3, Ki67, and CD31 staining of representative tumors harvested from mice. ** P
    Anti Cleaved Caspase 3, supplied by Servicebio Inc, used in various techniques. Bioz Stars score: 92/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Biocare Medical anti cleaved caspase 3
    TLR4 deficiency results in decreased neuronal apoptosis in the HF in response to GCI/R Neuronal apoptosis was confirmed by the methods of TUNEL staining (A to F) and IHC staining for cleaved <t>caspase-3,</t> a specific marker of apoptosis (G to L). The positive staining was shown in brown. Slides were counterstained with hematoxylin. Results show that GCI/R results in increased numbers of TUNEL positive cells in the HF (B and E) in WT mice. In TLR4 -/- -I/R mice, there were fewer TUNEL positive staining cells in the HF (C and F) compared with WT-I/R mice (M, # p
    Anti Cleaved Caspase 3, supplied by Biocare Medical, used in various techniques. Bioz Stars score: 92/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Epitomics anti cleaved caspase 3
    Protein expression in the kidney tissues using IHC staining and western blot analysis. IHC was used to determine the expression levels of HIF-1α ( a ), LC3 ( b ) and <t>cleaved-caspase-3</t> ( c ). d Western blot analysis of protein expression in kidney tissues. The mice were i.p. injected with ZnO NPs
    Anti Cleaved Caspase 3, supplied by Epitomics, used in various techniques. Bioz Stars score: 90/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti cleaved caspase 3
    Knockdown of endogenous BAIAP2L1 inhibits cell proliferation and enhances apoptosis induced by UV and cisplatin. BAIAP2L1 in two ovarian cancer cell lines (MDAH2774 and SKOV3) were knockdowned with si-RNA (A) and analyzed with (B) BrdU incorportation assay and (C) MTT assay. (D) BAIAP2L1 knockdown in SKOV3 cells increased cleavage of <t>caspase</t> 3 and PARP, which were indicators of apoptosis. The cells were irradiated with UV (100 J/M 2 ) or treated with 10 μM cisplatin for 24 h to induce apoptosis. To clearly visualize the differential intensities of cleaved PARP bands, two exposure times were used during chemiluminescence: long (60 sec) and short (10 sec). The actin level was used to normalize the amount of input protein. (E) Quantitative analyses of cleaved caspase 3 and PARP. Results shown are the mean ± standard error from three independent experiments. Asterisks denote statistical significance (p
    Anti Cleaved Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 11190 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti cleaved caspase 3
    miR-21 can reduce PC12 cell death and TRPM7 is a functional target of miR-21 (A) . MiR-21 transfection efficiency detected by qPCR (B) . Apoptosis rate of PC12 cells transfected with miR-21 mimics, miR-21 inhibitors, and their corresponding negative controls (NC) (C) . Fluo-4 assay of PC12 cells (D) . The expression level of TRPM7 in PC12 cells transfected with miR-21 mimics, miR-21 inhibitors, and their corresponding negative controls (NC) (E) . Target region of miR-21 and TRPM7 (F) . Luciferase assay for the combination of miR-21 and TRPM7 (G) . Western blot analyzing the expression level of TRPM7, p65, and p-IκB proteins (H) . Quantitative histogram of TRPM7 expression level in different groups (I) . Western blot analyzing the expression level of cleaved <t>caspase-3.</t> * p
    Anti Cleaved Caspase 3, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 818 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Synaptic Systems anti cleaved caspase 3
    Immunohistochemical co-localisation of cleaved <t>caspase</t> 3 with NeuN or GFAP. Brain sections tissue of 6 dpi SAFV infected or uninfected 2 week-old AG129 mice were stained with anti-cleaved caspase 3 and anti-GFAP or anti-NeuN. Resulting stain was viewed and taken with widefield or confocal fluorescence microscope. Cleaved caspase 3 is labelled in red, NeuN or GFAP is labelled in green, and DAPI is labelled in blue. Arrows point at examples of colocalisation
    Anti Cleaved Caspase 3, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 90/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    WuXi AppTec anti cleaved caspase 3 antibody
    Immunohistochemical co-localisation of cleaved <t>caspase</t> 3 with NeuN or GFAP. Brain sections tissue of 6 dpi SAFV infected or uninfected 2 week-old AG129 mice were stained with anti-cleaved caspase 3 and anti-GFAP or anti-NeuN. Resulting stain was viewed and taken with widefield or confocal fluorescence microscope. Cleaved caspase 3 is labelled in red, NeuN or GFAP is labelled in green, and DAPI is labelled in blue. Arrows point at examples of colocalisation
    Anti Cleaved Caspase 3 Antibody, supplied by WuXi AppTec, used in various techniques. Bioz Stars score: 93/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    SeP KO mice reduces apoptotic cells after myocardial I/R. LV tissue sections were subjected to TUNEL and 4′,6-diamidino-2-phenylindole (DAPI) staining. ( A ) Representative examples of TUNEL-positive myocytes in the ischemic area. Arrows indicate TUNEL-positive myocytes. ( B ) The number of TUNEL-positive myocytes was expressed as a percentage of total nuclei detected using DAPI staining. Heart homogenates were prepared from SeP KO and WT mice subjected to 30 min of ischemia and 24 h of reperfusion. Immunoblot analyses were performed using ( C ) anti-cleaved caspase-3 and GAPDH antibody. ( D ) Results of quantitative analysis of cleaved caspase-3 are shown. Data represent means from at least five mice each. * p

    Journal: International Journal of Molecular Sciences

    Article Title: Endogenous Selenoprotein P, a Liver-Derived Secretory Protein, Mediates Myocardial Ischemia/Reperfusion Injury in Mice

    doi: 10.3390/ijms19030878

    Figure Lengend Snippet: SeP KO mice reduces apoptotic cells after myocardial I/R. LV tissue sections were subjected to TUNEL and 4′,6-diamidino-2-phenylindole (DAPI) staining. ( A ) Representative examples of TUNEL-positive myocytes in the ischemic area. Arrows indicate TUNEL-positive myocytes. ( B ) The number of TUNEL-positive myocytes was expressed as a percentage of total nuclei detected using DAPI staining. Heart homogenates were prepared from SeP KO and WT mice subjected to 30 min of ischemia and 24 h of reperfusion. Immunoblot analyses were performed using ( C ) anti-cleaved caspase-3 and GAPDH antibody. ( D ) Results of quantitative analysis of cleaved caspase-3 are shown. Data represent means from at least five mice each. * p

    Article Snippet: The following antibodies were used for further analysis: anti-cleaved caspase-3 (#19677, Sigma-Aldrich, Tokyo, Japan), anti-phospho-IGF-I receptor β (#3024s, Cell Signaling, Tokyo, Japan), anti-Akt (#9272, Cell Signaling), anti-phospho-Akt (#4060, Cell Signaling), anti-p44/42 MAPK (Erk1/2) (#9102, Cell Signaling), anti-Phospho-p44/42 MAPK (Erk1/2) (#9101, Cell Signaling), anti-phospho-p70 S6 kinase (Thr421/Ser424) (#9204, Cell Signaling), and anti-GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA).

    Techniques: Mouse Assay, TUNEL Assay, Staining

    Effect of ALLO on the apoptotic index (cleaved CASP3 positive immunostained cells over the total number of cells) in follicles ( a ) and corpora lutea ( b ). Control groups follicles (1A upper panel), ALLO treated follicles (1A lower panel), control groups CL (1B upper panel), ALLO treated groups CL (1B lower panel); E, D1, D2, PE from left to right. Representative photomicrographs of follicles and corpora lutea immunostained for cleaved CASP3 at each stage of the estrous cycle. Values are expressed as mean ± S.E.M. Two-way ANOVA followed by Bonferroni’s post hoc, ( n =6), * p

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: Allopregnanolone alters follicular and luteal dynamics during the estrous cycle

    doi: 10.1186/s12958-018-0353-y

    Figure Lengend Snippet: Effect of ALLO on the apoptotic index (cleaved CASP3 positive immunostained cells over the total number of cells) in follicles ( a ) and corpora lutea ( b ). Control groups follicles (1A upper panel), ALLO treated follicles (1A lower panel), control groups CL (1B upper panel), ALLO treated groups CL (1B lower panel); E, D1, D2, PE from left to right. Representative photomicrographs of follicles and corpora lutea immunostained for cleaved CASP3 at each stage of the estrous cycle. Values are expressed as mean ± S.E.M. Two-way ANOVA followed by Bonferroni’s post hoc, ( n =6), * p

    Article Snippet: Anti-cleaved caspase 3 (CASP3) (Biocare Medical, #CP229C, California, USA) raised in rabbit, anti-proliferating cell nuclear antigen (PCNA) raised in rabbit (Santa Cruz Biotechnology, sc-7907, USA), anti-α-actin raised in mouse (Santa Cruz Biotechnology, sc-56499, USA), polyclonal anti-Von Willebrand factor raised in rabbit (Dako Cytomation, A0082, USA), Anti-Rabbit HRP IgG (Sigma Aldrich, A4914, USA), anti-mouse HRP IgG (R & D Systems HAF007, Minnesota, USA), and avidin-biotinylated HRP complex (Vectastain ABC system; Vector Laboratories, Burlingame, CA, USA) were used for immunohistochemistry technique.

    Techniques:

    BTLA signaling motifs show no effect on tumor killing, while Grb2 motif augments IL-2 production and T cell proliferation ( a ) Schematic diagram depicts the structure of BTLA WT (left), BTLAΔGrb2 (middle), and BTLAΔITSM (right). Signaling motifs with modified Tyrosine to Phenylalanine are indicated by a dotted pattern over a black background. A table summarizing the phenotype observed with the expression of BTLA or of the different BTLA constructs is presented on the right panel. ( b ) B16 OVA (mouse melanoma tumor positive for OVA) or B16F10 (mouse melanoma tumor negative for OVA) were stained with eFluor670® and co-cultured with OT-1 BTLA KO T cells overexpressing WT BTLA or BTLA mutants at the following T cell-to-tumor cell ratios (1:10, 1:3, and 1:1). Tumor cell death is depicted by the percentage of caspase-3 positive cells. N=3 independent experiments ( c ) OT-1 BTLA KO T cells overexpressing WT BTLA or its variants were re-stimulated with dendritic cells pulsed with OVA peptide. TNF-α and IFN-γ production by virally transduced OT-1 BTLA KO T cells was evaluated by intracellular staining. Bar graph depicts the percentage of positive cells (left panel) and mean fluorescence intensity (MFI) (right panel). Each bar represents three independent experiments. (Two-way ANOVA; * P

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: Multifaceted role of BTLA in the control of CD8+ T cell fate after antigen encounter

    doi: 10.1158/1078-0432.CCR-16-1217

    Figure Lengend Snippet: BTLA signaling motifs show no effect on tumor killing, while Grb2 motif augments IL-2 production and T cell proliferation ( a ) Schematic diagram depicts the structure of BTLA WT (left), BTLAΔGrb2 (middle), and BTLAΔITSM (right). Signaling motifs with modified Tyrosine to Phenylalanine are indicated by a dotted pattern over a black background. A table summarizing the phenotype observed with the expression of BTLA or of the different BTLA constructs is presented on the right panel. ( b ) B16 OVA (mouse melanoma tumor positive for OVA) or B16F10 (mouse melanoma tumor negative for OVA) were stained with eFluor670® and co-cultured with OT-1 BTLA KO T cells overexpressing WT BTLA or BTLA mutants at the following T cell-to-tumor cell ratios (1:10, 1:3, and 1:1). Tumor cell death is depicted by the percentage of caspase-3 positive cells. N=3 independent experiments ( c ) OT-1 BTLA KO T cells overexpressing WT BTLA or its variants were re-stimulated with dendritic cells pulsed with OVA peptide. TNF-α and IFN-γ production by virally transduced OT-1 BTLA KO T cells was evaluated by intracellular staining. Bar graph depicts the percentage of positive cells (left panel) and mean fluorescence intensity (MFI) (right panel). Each bar represents three independent experiments. (Two-way ANOVA; * P

    Article Snippet: After 3 h, the cells were fixed and permeabilized using BD Cytofix/Cytoperm (according to manufacturer protocol, BD Pharmingen™), then stained with anti-cleaved caspase-3 (clone CPP32, BD Pharmingen™), and analyzed by a BD FACSCanto II (BD Biosciences).

    Techniques: Modification, Expressing, Construct, Staining, Cell Culture, Fluorescence

    CD8 + BTLA + TIL demonstrate a superior anti-tumor effect in vivo ( a ) MART1-reactive CD8 + TIL (left and mid panel) and TIL 2549 (right panel) were sorted into BTLA + and BTLA − subsets. MEL 526 (melanoma tumor expressing MART-1 antigen, left and mid graph) or autologous tumor line 2549 (right graph) were stained with eFluor670® and co-cultured with TIL at the following TIL-to-tumor cell ratios (1:10, 1:3, and 1:1). Tumor cell death is measured by the percentage of caspase-3 positive cells. ( b) Ten million sorted CD8 + BTLA + or sorted CD8 + BTLA − TIL were intravenously injected into tumor bearing mice previously subcutaneously implanted with either MEL 526 or autologous melanoma tumor line 2549. Tumor burden was measured using calipers and diameter graphed as mm 2 . ( c ) Bar graph shows the percentage of CD45 + CD8 + in the peripheral blood on days 2, 4, 6, and 8 post-adoptive transfer in the same experiment described in ( b ). N= 5–8 animals per group. * P

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: Multifaceted role of BTLA in the control of CD8+ T cell fate after antigen encounter

    doi: 10.1158/1078-0432.CCR-16-1217

    Figure Lengend Snippet: CD8 + BTLA + TIL demonstrate a superior anti-tumor effect in vivo ( a ) MART1-reactive CD8 + TIL (left and mid panel) and TIL 2549 (right panel) were sorted into BTLA + and BTLA − subsets. MEL 526 (melanoma tumor expressing MART-1 antigen, left and mid graph) or autologous tumor line 2549 (right graph) were stained with eFluor670® and co-cultured with TIL at the following TIL-to-tumor cell ratios (1:10, 1:3, and 1:1). Tumor cell death is measured by the percentage of caspase-3 positive cells. ( b) Ten million sorted CD8 + BTLA + or sorted CD8 + BTLA − TIL were intravenously injected into tumor bearing mice previously subcutaneously implanted with either MEL 526 or autologous melanoma tumor line 2549. Tumor burden was measured using calipers and diameter graphed as mm 2 . ( c ) Bar graph shows the percentage of CD45 + CD8 + in the peripheral blood on days 2, 4, 6, and 8 post-adoptive transfer in the same experiment described in ( b ). N= 5–8 animals per group. * P

    Article Snippet: After 3 h, the cells were fixed and permeabilized using BD Cytofix/Cytoperm (according to manufacturer protocol, BD Pharmingen™), then stained with anti-cleaved caspase-3 (clone CPP32, BD Pharmingen™), and analyzed by a BD FACSCanto II (BD Biosciences).

    Techniques: In Vivo, Expressing, Staining, Cell Culture, Injection, Mouse Assay, Adoptive Transfer Assay

    Primary tumor growth is not affected by ABL kinase inhibitors MDA-MB 231/Dendra2 cells were injected into the mammary fat pad of 10-week-old SCID female mice and allowed to grow until the tumor reached the size of 100 mm 3 . At day 56 following injection, mice were treated by oral gavage with vehicle (5% DMSO, 2% hydroxypropyl cellulose, 0.5% Tween-80), 100 mg/kg imatinib, 70 mg/kg nilotinib or 100 mg/kg GNF-5 once a day, 5 days a week, for four weeks. ( A ) Time-dependent tumor growth. Tumor growth was assessed twice a week by measuring two perpendicular diameters and calculating tumor size in mm 3 . Treatment initiation is shown as red dotted line. n = 12 (vehicle), n = 10 (imatinib), n = 12 (nilotinib), n = 10 (GNF-5) mice per group from two independent experiments. ( B ) Primary tumors were dissected at the end of experiment and subjected to immunohistochemistry. Representative images of primary tumor sections stained with anti-PCNA (proliferation), anti-cleaved caspase 3 (apoptosis), and anti-CD31 (angiogenesis). ( C ) Quantification of PCNA positive cells (red/pink) normalized to DAPI positive cells (blue). The average percentage of proliferating cells in total cells per field is shown. ( D ) Quantification of apoptotic cells (cleaved caspase 3 positive cells). Shown is the number of apoptotic cells per field. ( E ) Quantification of CD-31 positive blood vessels. For all quantifications, n = 50 random fields from 5 tumors per condition. Bar, 100 µm.

    Journal: Oncotarget

    Article Title: Targeting invadopodia-mediated breast cancer metastasis by using ABL kinase inhibitors

    doi: 10.18632/oncotarget.25243

    Figure Lengend Snippet: Primary tumor growth is not affected by ABL kinase inhibitors MDA-MB 231/Dendra2 cells were injected into the mammary fat pad of 10-week-old SCID female mice and allowed to grow until the tumor reached the size of 100 mm 3 . At day 56 following injection, mice were treated by oral gavage with vehicle (5% DMSO, 2% hydroxypropyl cellulose, 0.5% Tween-80), 100 mg/kg imatinib, 70 mg/kg nilotinib or 100 mg/kg GNF-5 once a day, 5 days a week, for four weeks. ( A ) Time-dependent tumor growth. Tumor growth was assessed twice a week by measuring two perpendicular diameters and calculating tumor size in mm 3 . Treatment initiation is shown as red dotted line. n = 12 (vehicle), n = 10 (imatinib), n = 12 (nilotinib), n = 10 (GNF-5) mice per group from two independent experiments. ( B ) Primary tumors were dissected at the end of experiment and subjected to immunohistochemistry. Representative images of primary tumor sections stained with anti-PCNA (proliferation), anti-cleaved caspase 3 (apoptosis), and anti-CD31 (angiogenesis). ( C ) Quantification of PCNA positive cells (red/pink) normalized to DAPI positive cells (blue). The average percentage of proliferating cells in total cells per field is shown. ( D ) Quantification of apoptotic cells (cleaved caspase 3 positive cells). Shown is the number of apoptotic cells per field. ( E ) Quantification of CD-31 positive blood vessels. For all quantifications, n = 50 random fields from 5 tumors per condition. Bar, 100 µm.

    Article Snippet: Antibodies and reagents For immunofluorescence, anti-cortactin (ab-33333) and anti-PCNA were obtained from Abcam; anti-Arp2 (H-84) (SC-15389) and anti-Tks5 (FISH M-300) (SC-30122) were obtained from Santa Cruz Biotechnology; anti-pY421-cortactin (C0739) was obtained from Sigma-Aldrich; anti-CD31 was obtained from BD Biosciences; anti-cleaved caspase 3 was obtained from Cell Signaling Technology.

    Techniques: Multiple Displacement Amplification, Injection, Mouse Assay, Immunohistochemistry, Staining

    Expression of p53 is enhanced by miR-128 mimics via inhibition of MDM4 expression. (A) Western blot analysis of MDM4, p53 and cleaved caspase-3 expression following transfection with miR-128 mimics. (B) Quantification of MDM4 expression. (C) Quantification of p53 expression. (D) Quantification of cleaved caspase-3 expression. **P

    Journal: Experimental and Therapeutic Medicine

    Article Title: miR-128 induces pancreas cancer cell apoptosis by targeting MDM4

    doi: 10.3892/etm.2018.6047

    Figure Lengend Snippet: Expression of p53 is enhanced by miR-128 mimics via inhibition of MDM4 expression. (A) Western blot analysis of MDM4, p53 and cleaved caspase-3 expression following transfection with miR-128 mimics. (B) Quantification of MDM4 expression. (C) Quantification of p53 expression. (D) Quantification of cleaved caspase-3 expression. **P

    Article Snippet: The primary antibodies were as follows: Anti-p53 (cat. no. 2527, 1:300; Cell Signaling Technology, Inc., Danvers, MA, USA), anti-MDM4 (cat. no. sc-374147, 1:300; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), anti-cleaved caspase-3 (cat. no. sc-98785, 1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and anti-β-actin (cat. no. 7210, 1:1,000 Santa Cruz Biotechnology, Inc.).

    Techniques: Expressing, Inhibition, Western Blot, Transfection

    Percentage of live (Annexin V negative/PI negative cells), apoptotic (AnnexinV positive/PI negative cells) and dead (AnnexinV positive/PI positive cells or AnnexinV negative/PI positive cells) cells in LnCaP (A) and CW22rv1 (B) cell populations treated with continuous or pulsed Testosterone for 48 hours. The bars show the mean and SD of 36 replicates. Student-Newman-Keuls test for all pairwise comparisons in CW22rv1 and LnCaP cells at 48 hours after treatments. Expression levels, by western blot analysis, of pro-caspase-3 and cleaved caspase3 in LnCaP (C) and CW22rv1 (D) tumor cells tumor cells treated with continuous or pulsed Testosterone for 48 hours.

    Journal: Oncotarget

    Article Title: Episode-like pulse testosterone supplementation induces tumor senescence and growth arrest down-modulating androgen receptor through modulation of p-ERK1/2, pARser81 and CDK1 signaling: biological implications for men treated with testosterone replacement therapy

    doi: 10.18632/oncotarget.22776

    Figure Lengend Snippet: Percentage of live (Annexin V negative/PI negative cells), apoptotic (AnnexinV positive/PI negative cells) and dead (AnnexinV positive/PI positive cells or AnnexinV negative/PI positive cells) cells in LnCaP (A) and CW22rv1 (B) cell populations treated with continuous or pulsed Testosterone for 48 hours. The bars show the mean and SD of 36 replicates. Student-Newman-Keuls test for all pairwise comparisons in CW22rv1 and LnCaP cells at 48 hours after treatments. Expression levels, by western blot analysis, of pro-caspase-3 and cleaved caspase3 in LnCaP (C) and CW22rv1 (D) tumor cells tumor cells treated with continuous or pulsed Testosterone for 48 hours.

    Article Snippet: Anti-cyclin D1, anti-PCNA, anti-β-actin, anti-pro-caspsase-3, anti-cleaved caspase-3, anti-SKp2, anti-p27, anti-AR, anti-p-ERK1/2, anti-total ERK1/2, anti-NSE, anti-beclin-1, anti-CDK4 and p62 antibodies were purchased from SantaCruz (SantaCruz, CA).

    Techniques: Expressing, Western Blot

    P-MVs-secreted miR-191 affected tubular epithelial cell apoptosis via CBS. P-MVs isolated from patients with renal allograft rejection were divided into seven groups: control, P-MVs of healthy, P-MVs of patients, P-MVs of patients+NC, P-MVs of patients+miR-191 Inhibitor, P-MVs of patients+miR-191 inhibitor+si-control, P-MVs of patients+miR-191 Inhibitor+si-CBS. A. HK-2 cell apoptosis was observed by flow cytometry. B. Apoptosis related proteins of Bax, Bcl-2, Caspase 3 were measured. *P

    Journal: Cell Cycle

    Article Title: miR-191 secreted by platelet-derived microvesicles induced apoptosis of renal tubular epithelial cells and participated in renal ischemia-reperfusion injury via inhibiting CBS

    doi: 10.1080/15384101.2018.1542900

    Figure Lengend Snippet: P-MVs-secreted miR-191 affected tubular epithelial cell apoptosis via CBS. P-MVs isolated from patients with renal allograft rejection were divided into seven groups: control, P-MVs of healthy, P-MVs of patients, P-MVs of patients+NC, P-MVs of patients+miR-191 Inhibitor, P-MVs of patients+miR-191 inhibitor+si-control, P-MVs of patients+miR-191 Inhibitor+si-CBS. A. HK-2 cell apoptosis was observed by flow cytometry. B. Apoptosis related proteins of Bax, Bcl-2, Caspase 3 were measured. *P

    Article Snippet: Protein samples were isolated on 10% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride membranes (Invitrogen, USA) which were blocked in 5% skim milk for 2 h. Anti-CBS (Invitrogen), anti-cleaved caspase 3 (Merck Millipore), anti-BAX (Invitrogen), anti-Bcl2 (Invitrogen) and anti-β-Actin were used as the first primary antibody at 1:1000 dilutions.

    Techniques: Isolation, Flow Cytometry, Cytometry

    CBS expression, HK-2 cell apoptosis and apoptosis related proteins in allograft renal transplantation rat model. Rats were divided into four groups: sham+PBS, model+PBS, sham+miR-191 inhibitor (800 pmol), and model+miR-191 inhibitor (800 pmol). mRNA and protein levels of CBS (a), HK-2 cell apoptosis (b) apoptosis related proteins of Bax, Bcl-2, Caspase 3 (c) were measured. CBS expression, HK-2 cell apoptosis and apoptosis related proteins in renal autotransplantation rat model. Rats were divided into four groups: sham+PBS, model+PBS, sham+P-MVs (200 ug), and model+P-MVs (200 ug). mRNA and protein levels of CBS (d), HK-2 cell apoptosis (e) apoptosis related proteins of Bax, Bcl-2, Caspase 3 (f) were measured. *P

    Journal: Cell Cycle

    Article Title: miR-191 secreted by platelet-derived microvesicles induced apoptosis of renal tubular epithelial cells and participated in renal ischemia-reperfusion injury via inhibiting CBS

    doi: 10.1080/15384101.2018.1542900

    Figure Lengend Snippet: CBS expression, HK-2 cell apoptosis and apoptosis related proteins in allograft renal transplantation rat model. Rats were divided into four groups: sham+PBS, model+PBS, sham+miR-191 inhibitor (800 pmol), and model+miR-191 inhibitor (800 pmol). mRNA and protein levels of CBS (a), HK-2 cell apoptosis (b) apoptosis related proteins of Bax, Bcl-2, Caspase 3 (c) were measured. CBS expression, HK-2 cell apoptosis and apoptosis related proteins in renal autotransplantation rat model. Rats were divided into four groups: sham+PBS, model+PBS, sham+P-MVs (200 ug), and model+P-MVs (200 ug). mRNA and protein levels of CBS (d), HK-2 cell apoptosis (e) apoptosis related proteins of Bax, Bcl-2, Caspase 3 (f) were measured. *P

    Article Snippet: Protein samples were isolated on 10% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride membranes (Invitrogen, USA) which were blocked in 5% skim milk for 2 h. Anti-CBS (Invitrogen), anti-cleaved caspase 3 (Merck Millipore), anti-BAX (Invitrogen), anti-Bcl2 (Invitrogen) and anti-β-Actin were used as the first primary antibody at 1:1000 dilutions.

    Techniques: Expressing, Transplantation Assay

    Cells derived from Ela-myc pancreatic tumors are sensitive to sunitinib in vitro A. Analysis by Western blot of the expresssion of the sunitinib-targeted receptors VEGFR2, PDGFR-α and PDGFR–β in two different cell lines derived from Ela-myc pancreatic tumors. B. Bright field images showing Emyc-1 cell sensitivity to different doses of sunitinib (1, 2 or 4 μM) in vitro , as compared to cells treated with vehicle (DMSO). Scale bars, 100 μm. C. MTT experiments were performed to quantify cell viability upon sunitinib treatment (1, 2 or 4 μM) in the Emyc-1 cell line, observing a dose-dependent effect. D. Left, immunofluorescence of cleaved caspase 3, to detect cell apoptosis upon sunitinib treatment (1, 2 or 4 μM). Scale bars, 20 μm. Right, Western blot analysis of the levels of cleaved caspase 3 in Emyc-1 cell line treated with sunitinib. Tubulin levels are shown as the loading control. E. Left, Immunofluorescence of P-Histone H3, to show cell growth arrest upon sunitinib treatment (1, 2 or 4 μM) in Emyc-1 cells. Scale bars, 50 μm. Right, Bar plots showing quantification of P-Histone H3 immunofluorescence experiments on the right. * p

    Journal: Oncotarget

    Article Title: The pancreatic niche inhibits the effectiveness of sunitinib treatment of pancreatic cancer

    doi: 10.18632/oncotarget.10199

    Figure Lengend Snippet: Cells derived from Ela-myc pancreatic tumors are sensitive to sunitinib in vitro A. Analysis by Western blot of the expresssion of the sunitinib-targeted receptors VEGFR2, PDGFR-α and PDGFR–β in two different cell lines derived from Ela-myc pancreatic tumors. B. Bright field images showing Emyc-1 cell sensitivity to different doses of sunitinib (1, 2 or 4 μM) in vitro , as compared to cells treated with vehicle (DMSO). Scale bars, 100 μm. C. MTT experiments were performed to quantify cell viability upon sunitinib treatment (1, 2 or 4 μM) in the Emyc-1 cell line, observing a dose-dependent effect. D. Left, immunofluorescence of cleaved caspase 3, to detect cell apoptosis upon sunitinib treatment (1, 2 or 4 μM). Scale bars, 20 μm. Right, Western blot analysis of the levels of cleaved caspase 3 in Emyc-1 cell line treated with sunitinib. Tubulin levels are shown as the loading control. E. Left, Immunofluorescence of P-Histone H3, to show cell growth arrest upon sunitinib treatment (1, 2 or 4 μM) in Emyc-1 cells. Scale bars, 50 μm. Right, Bar plots showing quantification of P-Histone H3 immunofluorescence experiments on the right. * p

    Article Snippet: Antibodies used for specific tissue immunostaining included anti-VEGFR2 (Cell Signaling Technology), anti-PDGFR-α (R & D) anti-PDGFR-β (Cell Signaling Technology), anti-vWF (Neomarkers), anti-cleaved caspase 3 (R & D systems) anti-P-Histone H3 (Ser10) (Millipore) and anti-Ki67 (Novo Castra).

    Techniques: Derivative Assay, In Vitro, Western Blot, MTT Assay, Immunofluorescence

    Sunitinib effects in Ela-myc pancreatic tumor cell apoptosis and proliferation Tumor cell apoptosis and proliferation was analyzed for pancreatic tumors in control (Ctl) and sunitinib-treated Ela-myc mice. A. Tumor cell apoptosis was evaluated through cleaved caspase 3 (active form) staining. B. Tumor cell proliferation was evaluated through P-Histone H3 staining in acinar lesions, or Ki67 in ductal ones. Box-and-whisker plot quantifications are shown on the right (A, B). Scale bars, 50 μm.

    Journal: Oncotarget

    Article Title: The pancreatic niche inhibits the effectiveness of sunitinib treatment of pancreatic cancer

    doi: 10.18632/oncotarget.10199

    Figure Lengend Snippet: Sunitinib effects in Ela-myc pancreatic tumor cell apoptosis and proliferation Tumor cell apoptosis and proliferation was analyzed for pancreatic tumors in control (Ctl) and sunitinib-treated Ela-myc mice. A. Tumor cell apoptosis was evaluated through cleaved caspase 3 (active form) staining. B. Tumor cell proliferation was evaluated through P-Histone H3 staining in acinar lesions, or Ki67 in ductal ones. Box-and-whisker plot quantifications are shown on the right (A, B). Scale bars, 50 μm.

    Article Snippet: Antibodies used for specific tissue immunostaining included anti-VEGFR2 (Cell Signaling Technology), anti-PDGFR-α (R & D) anti-PDGFR-β (Cell Signaling Technology), anti-vWF (Neomarkers), anti-cleaved caspase 3 (R & D systems) anti-P-Histone H3 (Ser10) (Millipore) and anti-Ki67 (Novo Castra).

    Techniques: CTL Assay, Mouse Assay, Staining, Whisker Assay

    CRTH 2 deletion reduces doxorubicin‐induced cardiomyocyte apoptosis and cardiac injury in mice Western blot analysis of ER stress markers p‐IRE1 and cleaved ATF6 in mouse cardiomyocytes upon DOX (1 μmol/l) treatment. CRTH2 mRNA expression in mouse cardiomyocytes under DOX treatment. Data represent mean ± SEM. ** P = 0.00011, 12 h vs. 0 h; ** P = 0.000267, 24 h vs. 0 h (one‐way ANOVA); n = 4. Cardiac function in mice assessed by M‐mode echocardiographic analysis on day 7 after DOX treatment (20 mg/kg, i.p.). EF, ejection fraction (C); FS, fractional shortening (D); LVPWs, left ventricle posterior wall thickness at end‐systole (E). Data represent mean ± SEM. EF, * P = 0.00836, vs. WT; FS, * P = 0.00383, vs. WT; LVPWs, * P = 0.0198, vs. WT (unpaired two‐tailed t ‐test); WT, n = 15, CRTH2 −/− , n = 13. Kaplan–Meier survival curves for mice subjected to DOX treatment. * P = 0.0305, vs. WT (log‐rank test); n = 15. Representative TUNEL‐stained images of heart tissue in mice on day 7 after DOX treatment. Green, TUNEL‐positive nuclei; blue, DAPI; red, α‐actinin; scale bar, 50 μm. Quantification of TUNEL‐positive cardiomyocytes in (G). Data represent mean ± SEM. * P = 0.00278, vs. WT (unpaired two‐tailed t ‐test); n = 6. Western blot analysis of m‐calpain, caspase‐12, and caspase‐3 in heart tissue from DOX‐treated mice. Schematic diagram of CRTH2 promoting cardiomyocyte apoptosis under ER stress through the G αq /calpain/caspase‐12 signaling pathway. Source data are available online for this figure.

    Journal: EMBO Molecular Medicine

    Article Title: CRTH2 promotes endoplasmic reticulum stress‐induced cardiomyocyte apoptosis through m‐calpain

    doi: 10.15252/emmm.201708237

    Figure Lengend Snippet: CRTH 2 deletion reduces doxorubicin‐induced cardiomyocyte apoptosis and cardiac injury in mice Western blot analysis of ER stress markers p‐IRE1 and cleaved ATF6 in mouse cardiomyocytes upon DOX (1 μmol/l) treatment. CRTH2 mRNA expression in mouse cardiomyocytes under DOX treatment. Data represent mean ± SEM. ** P = 0.00011, 12 h vs. 0 h; ** P = 0.000267, 24 h vs. 0 h (one‐way ANOVA); n = 4. Cardiac function in mice assessed by M‐mode echocardiographic analysis on day 7 after DOX treatment (20 mg/kg, i.p.). EF, ejection fraction (C); FS, fractional shortening (D); LVPWs, left ventricle posterior wall thickness at end‐systole (E). Data represent mean ± SEM. EF, * P = 0.00836, vs. WT; FS, * P = 0.00383, vs. WT; LVPWs, * P = 0.0198, vs. WT (unpaired two‐tailed t ‐test); WT, n = 15, CRTH2 −/− , n = 13. Kaplan–Meier survival curves for mice subjected to DOX treatment. * P = 0.0305, vs. WT (log‐rank test); n = 15. Representative TUNEL‐stained images of heart tissue in mice on day 7 after DOX treatment. Green, TUNEL‐positive nuclei; blue, DAPI; red, α‐actinin; scale bar, 50 μm. Quantification of TUNEL‐positive cardiomyocytes in (G). Data represent mean ± SEM. * P = 0.00278, vs. WT (unpaired two‐tailed t ‐test); n = 6. Western blot analysis of m‐calpain, caspase‐12, and caspase‐3 in heart tissue from DOX‐treated mice. Schematic diagram of CRTH2 promoting cardiomyocyte apoptosis under ER stress through the G αq /calpain/caspase‐12 signaling pathway. Source data are available online for this figure.

    Article Snippet: Primary antibodies were diluted as follows: anti‐cleaved‐caspase‐3 (1:1,000; Signalway Antibody LLC), anti‐mouse GAPDH, anti‐HA‐tag, anti‐m‐calpain, anti‐μ‐calpain, anti‐caspase‐12, anti‐IRE1 (1:1,000; Cell Signaling Technology), anti‐calpain‐7 (1:1,000; ProteinTech Group, Chicago, IL, USA), anti‐P‐IRE1(1:1,000; Littleton, CO, USA), anti‐ATF6 (1:500, Santa Cruz, CA, USA), anti‐LC3A/B, anti‐phospho‐MLKL, anti‐MLKL (1:1,000; Cell Signaling Technology), anti‐caspase‐4, anti‐caspase‐8, and anti‐caspase‐9 (1:1,000; ABclonal).

    Techniques: Mouse Assay, Western Blot, Expressing, Two Tailed Test, TUNEL Assay, Staining

    CRTH 2 inhibition protects against myocardial infarction ( MI ) by reducing ischemia‐induced apoptosis in mice Representative TUNEL‐stained images of the peri‐infarct area in MI mouse heart. Green, TUNEL‐positive nuclei; blue, DAPI; red, α‐actinin; scale bar, 50 μm. Quantification of TUNEL‐positive cardiomyocytes. Data represent mean ± SEM. * P = 0.0147, vs. WT (unpaired two‐tailed t ‐test); WT, n = 6; CRTH2 −/− , n = 8. Western blot analysis of the activated caspase‐3 in the infarct border zone of mouse heart post‐MI. M‐mode echocardiographic analysis of cardiac function in mice at day 14 post‐MI. EF, ejection fraction (D); FS, fractional shortening (E); LVPWs, left ventricular posterior wall thickness at end‐systole. (F). Data represent mean ± SEM. EF, * P = 0.0015, vs. WT (unpaired two‐tailed t ‐test); FS, * P = 0.0015, vs. WT (unpaired two‐tailed t ‐test); LVPWs, * P = 0.00805, vs. WT (unpaired two‐tailed t ‐test); WT, n = 10; CRTH2 −/− , n = 12. Representative images of Evans blue and TTC‐stained mouse heart at day 14 post‐MI. Scale bar, 500 μm. Quantification of infarcted size in mouse heart after MI. Data represent mean ± SEM. ** P = 0.00354, vs. WT (unpaired two‐tailed t ‐test); n = 10. Heart weight‐to‐body weight ratio in mice subjected to MI. Data represent mean ± SEM. * P = 0.0119, vs. WT (unpaired two‐tailed t ‐test); WT and CRTH2 −/− (Sham), n = 8; WT and CRTH2 −/− (MI), n = 10. Kaplan–Meier survival curves for mice at day 14 post‐MI. * P = 0.0356, vs. WT (log‐rank test).WT, n = 29; CRTH2 −/− , n = 26. Representative images of Masson's trichrome staining of cardiac tissues from infarcted hearts. Scale bar, 20 μm. Quantification of collagen content in (K). Data represent mean ± SEM. * P = 0.0196, vs. WT (unpaired two‐tailed t ‐test); WT and CRTH2 −/− (Sham), n = 7; WT and CRTH2 −/− (MI), n = 9. Source data are available online for this figure.

    Journal: EMBO Molecular Medicine

    Article Title: CRTH2 promotes endoplasmic reticulum stress‐induced cardiomyocyte apoptosis through m‐calpain

    doi: 10.15252/emmm.201708237

    Figure Lengend Snippet: CRTH 2 inhibition protects against myocardial infarction ( MI ) by reducing ischemia‐induced apoptosis in mice Representative TUNEL‐stained images of the peri‐infarct area in MI mouse heart. Green, TUNEL‐positive nuclei; blue, DAPI; red, α‐actinin; scale bar, 50 μm. Quantification of TUNEL‐positive cardiomyocytes. Data represent mean ± SEM. * P = 0.0147, vs. WT (unpaired two‐tailed t ‐test); WT, n = 6; CRTH2 −/− , n = 8. Western blot analysis of the activated caspase‐3 in the infarct border zone of mouse heart post‐MI. M‐mode echocardiographic analysis of cardiac function in mice at day 14 post‐MI. EF, ejection fraction (D); FS, fractional shortening (E); LVPWs, left ventricular posterior wall thickness at end‐systole. (F). Data represent mean ± SEM. EF, * P = 0.0015, vs. WT (unpaired two‐tailed t ‐test); FS, * P = 0.0015, vs. WT (unpaired two‐tailed t ‐test); LVPWs, * P = 0.00805, vs. WT (unpaired two‐tailed t ‐test); WT, n = 10; CRTH2 −/− , n = 12. Representative images of Evans blue and TTC‐stained mouse heart at day 14 post‐MI. Scale bar, 500 μm. Quantification of infarcted size in mouse heart after MI. Data represent mean ± SEM. ** P = 0.00354, vs. WT (unpaired two‐tailed t ‐test); n = 10. Heart weight‐to‐body weight ratio in mice subjected to MI. Data represent mean ± SEM. * P = 0.0119, vs. WT (unpaired two‐tailed t ‐test); WT and CRTH2 −/− (Sham), n = 8; WT and CRTH2 −/− (MI), n = 10. Kaplan–Meier survival curves for mice at day 14 post‐MI. * P = 0.0356, vs. WT (log‐rank test).WT, n = 29; CRTH2 −/− , n = 26. Representative images of Masson's trichrome staining of cardiac tissues from infarcted hearts. Scale bar, 20 μm. Quantification of collagen content in (K). Data represent mean ± SEM. * P = 0.0196, vs. WT (unpaired two‐tailed t ‐test); WT and CRTH2 −/− (Sham), n = 7; WT and CRTH2 −/− (MI), n = 9. Source data are available online for this figure.

    Article Snippet: Primary antibodies were diluted as follows: anti‐cleaved‐caspase‐3 (1:1,000; Signalway Antibody LLC), anti‐mouse GAPDH, anti‐HA‐tag, anti‐m‐calpain, anti‐μ‐calpain, anti‐caspase‐12, anti‐IRE1 (1:1,000; Cell Signaling Technology), anti‐calpain‐7 (1:1,000; ProteinTech Group, Chicago, IL, USA), anti‐P‐IRE1(1:1,000; Littleton, CO, USA), anti‐ATF6 (1:500, Santa Cruz, CA, USA), anti‐LC3A/B, anti‐phospho‐MLKL, anti‐MLKL (1:1,000; Cell Signaling Technology), anti‐caspase‐4, anti‐caspase‐8, and anti‐caspase‐9 (1:1,000; ABclonal).

    Techniques: Inhibition, Mouse Assay, TUNEL Assay, Staining, Two Tailed Test, Western Blot

    CRTH 2 deletion attenuates anoxia‐induced cardiomyocyte apoptosis in vitro Representative TUNEL‐stained images of mouse cardiomyocytes challenged with anoxia. Green, TUNEL‐positive nuclei; blue, DAPI‐stained nuclei; red, cardiomyocytes labeled with antibody to α‐actinin; scale bar, 50 μm. Quantification of TUNEL‐positive cells in (A). Data represent mean ± SEM. **P = 0.00202, vs. WT (Mann–Whitney U ‐test); n = 6. Flow cytometry analysis of annexin V and propidium iodide (PI) staining of apoptotic cardiomyocytes following anoxia treatment. Quantification of annexin V + cells in (C). Data represent mean ± SEM. * P = 0.0002, vs. WT (unpaired two‐tailed t ‐test); n = 6. Western blot analysis of activated caspase‐3 in mouse cardiomyocytes challenged with anoxia. Source data are available online for this figure.

    Journal: EMBO Molecular Medicine

    Article Title: CRTH2 promotes endoplasmic reticulum stress‐induced cardiomyocyte apoptosis through m‐calpain

    doi: 10.15252/emmm.201708237

    Figure Lengend Snippet: CRTH 2 deletion attenuates anoxia‐induced cardiomyocyte apoptosis in vitro Representative TUNEL‐stained images of mouse cardiomyocytes challenged with anoxia. Green, TUNEL‐positive nuclei; blue, DAPI‐stained nuclei; red, cardiomyocytes labeled with antibody to α‐actinin; scale bar, 50 μm. Quantification of TUNEL‐positive cells in (A). Data represent mean ± SEM. **P = 0.00202, vs. WT (Mann–Whitney U ‐test); n = 6. Flow cytometry analysis of annexin V and propidium iodide (PI) staining of apoptotic cardiomyocytes following anoxia treatment. Quantification of annexin V + cells in (C). Data represent mean ± SEM. * P = 0.0002, vs. WT (unpaired two‐tailed t ‐test); n = 6. Western blot analysis of activated caspase‐3 in mouse cardiomyocytes challenged with anoxia. Source data are available online for this figure.

    Article Snippet: Primary antibodies were diluted as follows: anti‐cleaved‐caspase‐3 (1:1,000; Signalway Antibody LLC), anti‐mouse GAPDH, anti‐HA‐tag, anti‐m‐calpain, anti‐μ‐calpain, anti‐caspase‐12, anti‐IRE1 (1:1,000; Cell Signaling Technology), anti‐calpain‐7 (1:1,000; ProteinTech Group, Chicago, IL, USA), anti‐P‐IRE1(1:1,000; Littleton, CO, USA), anti‐ATF6 (1:500, Santa Cruz, CA, USA), anti‐LC3A/B, anti‐phospho‐MLKL, anti‐MLKL (1:1,000; Cell Signaling Technology), anti‐caspase‐4, anti‐caspase‐8, and anti‐caspase‐9 (1:1,000; ABclonal).

    Techniques: In Vitro, TUNEL Assay, Staining, Labeling, MANN-WHITNEY, Flow Cytometry, Cytometry, Two Tailed Test, Western Blot

    Effects of PCNP on the apoptosis of human neuroblastoma cells. a The apoptotic levels of SH-SY5Y and SK-N-SH cells were measured by TUNEL staining; original magnification 100×. b The percentages of TUNEL-positive cells were calculated by the formula shown above. c Western blotting analysis for the expression of cleaved caspase-3, − 8, and − 9 and cleaved PARP in SH-SY5Y and SK-N-SH cells. GAPDH was used as the loading control. ( d, e ) The densitometry analysis of each factor was performed in SH-SY5Y and SK-N-SH cells, normalized to the corresponding GAPDH level. Data are presented as mean ± SEM of three independent experiments; * P

    Journal: BMC Cancer

    Article Title: PEST-containing nuclear protein mediates the proliferation, migration, and invasion of human neuroblastoma cells through MAPK and PI3K/AKT/mTOR signaling pathways

    doi: 10.1186/s12885-018-4391-9

    Figure Lengend Snippet: Effects of PCNP on the apoptosis of human neuroblastoma cells. a The apoptotic levels of SH-SY5Y and SK-N-SH cells were measured by TUNEL staining; original magnification 100×. b The percentages of TUNEL-positive cells were calculated by the formula shown above. c Western blotting analysis for the expression of cleaved caspase-3, − 8, and − 9 and cleaved PARP in SH-SY5Y and SK-N-SH cells. GAPDH was used as the loading control. ( d, e ) The densitometry analysis of each factor was performed in SH-SY5Y and SK-N-SH cells, normalized to the corresponding GAPDH level. Data are presented as mean ± SEM of three independent experiments; * P

    Article Snippet: Anti-PCNP, Anti-B-cell lymphoma-2 (Bcl-2), anti-Bcl-2-associated X protein (Bax), anti-B-cell lymphoma-extra large (Bcl-xl), anti-Bcl-xl/Bcl-2-associated death promoter (Bad), anti-cleaved caspase-3, anti-cleaved caspase-8, anti-cleaved caspase-9, anti-Cleaved poly adenosine diphosphate-ribose polymerase (PARP), and anti-GAPDH antibodies were purchased from ProteinTech (Chicago, IL, USA).

    Techniques: TUNEL Assay, Staining, Western Blot, Expressing

    SCIN knockdown activated caspase 3 signalling pathway and induced cytochrome c .

    Journal: FEBS Open Bio

    Article Title: Loss of scinderin decreased expression of epidermal growth factor receptor and promoted apoptosis of castration‐resistant prostate cancer cells

    doi: 10.1002/2211-5463.12412

    Figure Lengend Snippet: SCIN knockdown activated caspase 3 signalling pathway and induced cytochrome c .

    Article Snippet: After blocking the membrane with 5% non‐fat milk, target proteins were detected using the following antibodies: anti‐SCIN, anti‐epidermal growth factor receptor (EGFR) anti‐B‐cell lymphoma‐extra‐large (Bcl‐xl), anti‐caspase 9, anti‐cleaved caspase 3, anti‐cytochrome c (all 1 : 1000; Proteintech, Rosemont, IL, USA), anti‐ glyceraldehyde 3‐phosphate dehydrogenase (GAPDH, 1 : 500 000; all Proteintech), anti‐mitogen‐activated protein kinase kinase (MEK), anti‐phosphorylated (p)‐MEK (both 1 : 1000; SAB, College Park, MD, USA), anti‐extracellular signal‐regulated kinase (ERK), anti‐p‐ERK (both 1 : 3000; Santa Cruz Biotechnology, Dallas, TX, USA), and anti‐Akt (1 : 1000; Cell Signaling Technology, Danvers, MA, USA), anti‐p‐AKT, anti‐poly‐ADP ribose polymerase (PARP; both 1 : 1000; Cell Signaling Technology).

    Techniques:

    Oligodendrocyte precursor cell susceptibility to apoptotic induction following inflammatory stress. Analysis of Casp3 expression in O4+ cells following an inflammatory stress. The fold change of apoptotic (Casp3+) and viable (Casp3-) O4+ cells was similar in the WT and Hp70.1 KO CNS cell cultures after 24 h of stimulation with LPS plus IFN-γ. The error bars correspond to the SEM.

    Journal: PLoS ONE

    Article Title: Hsp70 Regulates Immune Response in Experimental Autoimmune Encephalomyelitis

    doi: 10.1371/journal.pone.0105737

    Figure Lengend Snippet: Oligodendrocyte precursor cell susceptibility to apoptotic induction following inflammatory stress. Analysis of Casp3 expression in O4+ cells following an inflammatory stress. The fold change of apoptotic (Casp3+) and viable (Casp3-) O4+ cells was similar in the WT and Hp70.1 KO CNS cell cultures after 24 h of stimulation with LPS plus IFN-γ. The error bars correspond to the SEM.

    Article Snippet: For immunostaining of the CNS cultures, the following primary antibodies were used: anti-O4 (OPC) (mouse IgM, 1∶5, hybridoma generously donated by Dr. Sommer), anti-myelin basic protein (MBP, mature oligodendrocytes) (chicken IgY, 1∶200, Millipore), anti-cleaved Caspase 3 (Casp3) (rabbit polyclonal, 1∶500, Cell Signaling Technologies, Danvers, MA, USA), anti-NeuN (neurons) (mouse IgG1, 1∶300, Merck Millipore), anti-GFAP (astrocytes) (rabbit polyclonal, 1∶500, Dako), isolectin B4 conjugated to fluorescein-isothiocyanate (FITC) (mouse microglial cells) (1∶1000, Sigma), anti-CD11b/c (rat microglial cells) (mouse IgG2a, 1∶100, BD Pharmingen, San Diego, CA) and anti-CD68 (activated rat microglial cells) (mouse IgG1 1∶200, AbD Serotec, UK).

    Techniques: Expressing

    Apoptotic induction in CNS cell cultures. Apoptotic immunocytochemical detection in oligodendrocyte precursor cells (OPCs, O4+ cells) and mature oligodendrocytes (MBP+ cells) in mixed CNS cell cultures of WT and Hsp70.1 KO mice under non-stimulated (left column) and LPS plus IFN-γ-stimulated (right column) culture conditions. A. Detection of apoptotic OPCs cells (O4+Casp3+ cells). The rows indicate the O4+Casp3+ cells. B. Detection of apoptotic mature oligodendrocytes (MBP+Casp3+ cells).

    Journal: PLoS ONE

    Article Title: Hsp70 Regulates Immune Response in Experimental Autoimmune Encephalomyelitis

    doi: 10.1371/journal.pone.0105737

    Figure Lengend Snippet: Apoptotic induction in CNS cell cultures. Apoptotic immunocytochemical detection in oligodendrocyte precursor cells (OPCs, O4+ cells) and mature oligodendrocytes (MBP+ cells) in mixed CNS cell cultures of WT and Hsp70.1 KO mice under non-stimulated (left column) and LPS plus IFN-γ-stimulated (right column) culture conditions. A. Detection of apoptotic OPCs cells (O4+Casp3+ cells). The rows indicate the O4+Casp3+ cells. B. Detection of apoptotic mature oligodendrocytes (MBP+Casp3+ cells).

    Article Snippet: For immunostaining of the CNS cultures, the following primary antibodies were used: anti-O4 (OPC) (mouse IgM, 1∶5, hybridoma generously donated by Dr. Sommer), anti-myelin basic protein (MBP, mature oligodendrocytes) (chicken IgY, 1∶200, Millipore), anti-cleaved Caspase 3 (Casp3) (rabbit polyclonal, 1∶500, Cell Signaling Technologies, Danvers, MA, USA), anti-NeuN (neurons) (mouse IgG1, 1∶300, Merck Millipore), anti-GFAP (astrocytes) (rabbit polyclonal, 1∶500, Dako), isolectin B4 conjugated to fluorescein-isothiocyanate (FITC) (mouse microglial cells) (1∶1000, Sigma), anti-CD11b/c (rat microglial cells) (mouse IgG2a, 1∶100, BD Pharmingen, San Diego, CA) and anti-CD68 (activated rat microglial cells) (mouse IgG1 1∶200, AbD Serotec, UK).

    Techniques: Mouse Assay

    Upregulating H3K27me3 levels sensitizes OS to cisplatin in vivo.  a  Macroscopic image of tumor size after mice were sacrificed. Tumor volume ( b ) and weight ( c ) were measured at the indicated times.  d  H  E, TUNEL, cleaved Caspase 3, Ki67, and CD31 staining of representative tumors harvested from mice. ** P

    Journal: Clinical Epigenetics

    Article Title: Elevated H3K27me3 levels sensitize osteosarcoma to cisplatin

    doi: 10.1186/s13148-018-0605-x

    Figure Lengend Snippet: Upregulating H3K27me3 levels sensitizes OS to cisplatin in vivo. a Macroscopic image of tumor size after mice were sacrificed. Tumor volume ( b ) and weight ( c ) were measured at the indicated times. d H E, TUNEL, cleaved Caspase 3, Ki67, and CD31 staining of representative tumors harvested from mice. ** P

    Article Snippet: After deparaffinization, rehydration, antigen retrieval, and blockade of endogenous peroxidase, tissues were incubated with anti-TUNEL (Roche), anti-cleaved Caspase 3 (Servicebio), anti-Ki67 (Servicebio), and anti-CD31 (Abways) primary antibodies at 4 °C overnight.

    Techniques: In Vivo, Mouse Assay, TUNEL Assay, Staining

    Upregulating H3K27me3 levels sensitizes OS to cisplatin in vivo.  a  Macroscopic image of tumor size after mice were sacrificed. Tumor volume ( b ) and weight ( c ) were measured at the indicated times.  d  H  E, TUNEL, cleaved Caspase 3, Ki67, and CD31 staining of representative tumors harvested from mice. ** P

    Journal: Clinical Epigenetics

    Article Title: Elevated H3K27me3 levels sensitize osteosarcoma to cisplatin

    doi: 10.1186/s13148-018-0605-x

    Figure Lengend Snippet: Upregulating H3K27me3 levels sensitizes OS to cisplatin in vivo. a Macroscopic image of tumor size after mice were sacrificed. Tumor volume ( b ) and weight ( c ) were measured at the indicated times. d H E, TUNEL, cleaved Caspase 3, Ki67, and CD31 staining of representative tumors harvested from mice. ** P

    Article Snippet: Immunohistochemistry After deparaffinization, rehydration, antigen retrieval, and blockade of endogenous peroxidase, tissues were incubated with anti-TUNEL (Roche), anti-cleaved Caspase 3 (Servicebio), anti-Ki67 (Servicebio), and anti-CD31 (Abways) primary antibodies at 4 °C overnight.

    Techniques: In Vivo, Mouse Assay, TUNEL Assay, Staining

    TLR4 deficiency results in decreased neuronal apoptosis in the HF in response to GCI/R Neuronal apoptosis was confirmed by the methods of TUNEL staining (A to F) and IHC staining for cleaved caspase-3, a specific marker of apoptosis (G to L). The positive staining was shown in brown. Slides were counterstained with hematoxylin. Results show that GCI/R results in increased numbers of TUNEL positive cells in the HF (B and E) in WT mice. In TLR4 -/- -I/R mice, there were fewer TUNEL positive staining cells in the HF (C and F) compared with WT-I/R mice (M, # p

    Journal: Journal of neuroimmunology

    Article Title: Activation of Toll-like Receptor 4 Signaling Contributes to Hippocampal Neuronal Death Following Global Cerebral Ischemia/Reperfusion

    doi: 10.1016/j.jneuroim.2007.08.014

    Figure Lengend Snippet: TLR4 deficiency results in decreased neuronal apoptosis in the HF in response to GCI/R Neuronal apoptosis was confirmed by the methods of TUNEL staining (A to F) and IHC staining for cleaved caspase-3, a specific marker of apoptosis (G to L). The positive staining was shown in brown. Slides were counterstained with hematoxylin. Results show that GCI/R results in increased numbers of TUNEL positive cells in the HF (B and E) in WT mice. In TLR4 -/- -I/R mice, there were fewer TUNEL positive staining cells in the HF (C and F) compared with WT-I/R mice (M, # p

    Article Snippet: The primary antibodies used in the present study were anti-TLR4 (1:1000, gift from R. Medzhitov, Yale University), anti-p-NFκB-p65 (1:50, #3037, Cell Signaling Technology, Inc.) and anti-cleaved caspase-3 (1:50, CP229, Biocare Medical, Concord CA 94520).

    Techniques: TUNEL Assay, Staining, Immunohistochemistry, Marker, Mouse Assay

    Protein expression in the kidney tissues using IHC staining and western blot analysis. IHC was used to determine the expression levels of HIF-1α ( a ), LC3 ( b ) and cleaved-caspase-3 ( c ). d Western blot analysis of protein expression in kidney tissues. The mice were i.p. injected with ZnO NPs

    Journal: Particle and Fibre Toxicology

    Article Title: The role of hypoxia-inducible factor-1α in zinc oxide nanoparticle-induced nephrotoxicity in vitro and in vivo

    doi: 10.1186/s12989-016-0163-3

    Figure Lengend Snippet: Protein expression in the kidney tissues using IHC staining and western blot analysis. IHC was used to determine the expression levels of HIF-1α ( a ), LC3 ( b ) and cleaved-caspase-3 ( c ). d Western blot analysis of protein expression in kidney tissues. The mice were i.p. injected with ZnO NPs

    Article Snippet: Tissue sections were immunohistochemically incubated overnight at 4 °C with the anti-LC3 (MBL, Japan), anti-HIF-1α (BD Transduction Laboratories, USA) or anti-cleaved-caspase 3 (Epitomics, USA) antibodies.

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

    Knockdown of endogenous BAIAP2L1 inhibits cell proliferation and enhances apoptosis induced by UV and cisplatin. BAIAP2L1 in two ovarian cancer cell lines (MDAH2774 and SKOV3) were knockdowned with si-RNA (A) and analyzed with (B) BrdU incorportation assay and (C) MTT assay. (D) BAIAP2L1 knockdown in SKOV3 cells increased cleavage of caspase 3 and PARP, which were indicators of apoptosis. The cells were irradiated with UV (100 J/M 2 ) or treated with 10 μM cisplatin for 24 h to induce apoptosis. To clearly visualize the differential intensities of cleaved PARP bands, two exposure times were used during chemiluminescence: long (60 sec) and short (10 sec). The actin level was used to normalize the amount of input protein. (E) Quantitative analyses of cleaved caspase 3 and PARP. Results shown are the mean ± standard error from three independent experiments. Asterisks denote statistical significance (p

    Journal: PLoS ONE

    Article Title: BAI1-Associated Protein 2-Like 1 (BAIAP2L1) Is a Potential Biomarker in Ovarian Cancer

    doi: 10.1371/journal.pone.0133081

    Figure Lengend Snippet: Knockdown of endogenous BAIAP2L1 inhibits cell proliferation and enhances apoptosis induced by UV and cisplatin. BAIAP2L1 in two ovarian cancer cell lines (MDAH2774 and SKOV3) were knockdowned with si-RNA (A) and analyzed with (B) BrdU incorportation assay and (C) MTT assay. (D) BAIAP2L1 knockdown in SKOV3 cells increased cleavage of caspase 3 and PARP, which were indicators of apoptosis. The cells were irradiated with UV (100 J/M 2 ) or treated with 10 μM cisplatin for 24 h to induce apoptosis. To clearly visualize the differential intensities of cleaved PARP bands, two exposure times were used during chemiluminescence: long (60 sec) and short (10 sec). The actin level was used to normalize the amount of input protein. (E) Quantitative analyses of cleaved caspase 3 and PARP. Results shown are the mean ± standard error from three independent experiments. Asterisks denote statistical significance (p

    Article Snippet: Protein samples were analyzed using anti-BAIAP2L1 (Abnova, Taipei, Taiwan), anti-cleaved caspase 3 (Cell Signaling Technology, Danvers, MA, USA), anti-PARP (Cell Signaling Technology, Danvers, MA, USA) and anti-actin (Santa Cruz Biotechnology, Santa Cruz, CA) as primary antibodies, and corresponding horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnology, Santa Cruz, CA).

    Techniques: MTT Assay, Irradiation, Size-exclusion Chromatography

    miR-21 can reduce PC12 cell death and TRPM7 is a functional target of miR-21 (A) . MiR-21 transfection efficiency detected by qPCR (B) . Apoptosis rate of PC12 cells transfected with miR-21 mimics, miR-21 inhibitors, and their corresponding negative controls (NC) (C) . Fluo-4 assay of PC12 cells (D) . The expression level of TRPM7 in PC12 cells transfected with miR-21 mimics, miR-21 inhibitors, and their corresponding negative controls (NC) (E) . Target region of miR-21 and TRPM7 (F) . Luciferase assay for the combination of miR-21 and TRPM7 (G) . Western blot analyzing the expression level of TRPM7, p65, and p-IκB proteins (H) . Quantitative histogram of TRPM7 expression level in different groups (I) . Western blot analyzing the expression level of cleaved caspase-3. * p

    Journal: Frontiers in Neurology

    Article Title: MicroRNA-21 Overexpression Promotes the Neuroprotective Efficacy of Mesenchymal Stem Cells for Treatment of Intracerebral Hemorrhage

    doi: 10.3389/fneur.2018.00931

    Figure Lengend Snippet: miR-21 can reduce PC12 cell death and TRPM7 is a functional target of miR-21 (A) . MiR-21 transfection efficiency detected by qPCR (B) . Apoptosis rate of PC12 cells transfected with miR-21 mimics, miR-21 inhibitors, and their corresponding negative controls (NC) (C) . Fluo-4 assay of PC12 cells (D) . The expression level of TRPM7 in PC12 cells transfected with miR-21 mimics, miR-21 inhibitors, and their corresponding negative controls (NC) (E) . Target region of miR-21 and TRPM7 (F) . Luciferase assay for the combination of miR-21 and TRPM7 (G) . Western blot analyzing the expression level of TRPM7, p65, and p-IκB proteins (H) . Quantitative histogram of TRPM7 expression level in different groups (I) . Western blot analyzing the expression level of cleaved caspase-3. * p

    Article Snippet: The membranes were incubated in 1% BSA solution with primary antibodies including anti-cleaved caspase-3, anti-matrix metalloproteinase 2 (MMP2), anti-matrix metalloproteinase 9 (MMP9), anti-tissue inhibitor of metalloproteinase-1 (TIMP-1)TIMP1, anti-transient receptor potential melastatin 7 (TRPM7), anti-p65, anti-phospho-IkB, and anti-beta actin (Abcam, CA, United States) at 4°C overnight and then incubated with the secondary antibody for 1 h at 22–25°C.

    Techniques: Functional Assay, Transfection, Real-time Polymerase Chain Reaction, Expressing, Luciferase, Western Blot

    miR-21 enhanced survival of Hemin-induced mesenchymal stem cells (MSCs) (A) . Viability of MSCs treated with Hemin for different lengths of time (B) . Expression level of miR-21 detected by qPCR (C) . Viability of MSCs transfected with miR-21 mimics, miR-21 inhibitors, or their corresponding negative control (NC) (D) . Cell apoptosis analyzed by Annexin V-FITC and PI staining of MSCs in different groups (E) . Western blot analysis of cleaved-caspase-3 expression level in MSCs transfected with different miRNAs. * p

    Journal: Frontiers in Neurology

    Article Title: MicroRNA-21 Overexpression Promotes the Neuroprotective Efficacy of Mesenchymal Stem Cells for Treatment of Intracerebral Hemorrhage

    doi: 10.3389/fneur.2018.00931

    Figure Lengend Snippet: miR-21 enhanced survival of Hemin-induced mesenchymal stem cells (MSCs) (A) . Viability of MSCs treated with Hemin for different lengths of time (B) . Expression level of miR-21 detected by qPCR (C) . Viability of MSCs transfected with miR-21 mimics, miR-21 inhibitors, or their corresponding negative control (NC) (D) . Cell apoptosis analyzed by Annexin V-FITC and PI staining of MSCs in different groups (E) . Western blot analysis of cleaved-caspase-3 expression level in MSCs transfected with different miRNAs. * p

    Article Snippet: The membranes were incubated in 1% BSA solution with primary antibodies including anti-cleaved caspase-3, anti-matrix metalloproteinase 2 (MMP2), anti-matrix metalloproteinase 9 (MMP9), anti-tissue inhibitor of metalloproteinase-1 (TIMP-1)TIMP1, anti-transient receptor potential melastatin 7 (TRPM7), anti-p65, anti-phospho-IkB, and anti-beta actin (Abcam, CA, United States) at 4°C overnight and then incubated with the secondary antibody for 1 h at 22–25°C.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Transfection, Negative Control, Staining, Western Blot

    Immunohistochemical co-localisation of cleaved caspase 3 with NeuN or GFAP. Brain sections tissue of 6 dpi SAFV infected or uninfected 2 week-old AG129 mice were stained with anti-cleaved caspase 3 and anti-GFAP or anti-NeuN. Resulting stain was viewed and taken with widefield or confocal fluorescence microscope. Cleaved caspase 3 is labelled in red, NeuN or GFAP is labelled in green, and DAPI is labelled in blue. Arrows point at examples of colocalisation

    Journal: Virology Journal

    Article Title: Immunohistochemical insights into Saffold virus infection of the brain of juvenile AG129 mice

    doi: 10.1186/s12985-016-0654-8

    Figure Lengend Snippet: Immunohistochemical co-localisation of cleaved caspase 3 with NeuN or GFAP. Brain sections tissue of 6 dpi SAFV infected or uninfected 2 week-old AG129 mice were stained with anti-cleaved caspase 3 and anti-GFAP or anti-NeuN. Resulting stain was viewed and taken with widefield or confocal fluorescence microscope. Cleaved caspase 3 is labelled in red, NeuN or GFAP is labelled in green, and DAPI is labelled in blue. Arrows point at examples of colocalisation

    Article Snippet: The same immunohistochemistry protocol as above was used, except primary antibodies used were anti-cleaved caspase 3 (Synaptic Systems) and guinea pig anti-GFAP or guinea pig anti-NeuN (Synaptic Systems).

    Techniques: Immunohistochemistry, Infection, Mouse Assay, Staining, Fluorescence, Microscopy