cleaved caspase-3 Search Results


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  • 98
    Millipore cleaved caspase 3
    Generation of 3D-spheroid GMSCs. ( A ) 4 × 10 4 of GMSCs/well in 200 µl of complete stemgro culture medium was seeded into each well of ultra-low attachment round-shaped 96-U well plates and cultured for 48 h. H E staining of cryosections of GMSC spheroids. Scale bar: 50 µm. ( B ) Immunocytochemistry showed the expression of a panel of MSC-associated markers CD29, CD73 and CD90 and extracellular components such as type I collagen (col-I), vimentin, fibronectin, and laminin in GMSC-derived spheroids. ( C ) GMSC spheroids were dissociated into single cells and stained with Annexin V-FITC and 7-AAD to detect early apoptosis and late apoptotic/necrotic cells by flow cytometric analysis. ( D ) Immunocytochemistry showed that less than 10% of cells inside GMSC spheroids were positive for the cleaved <t>caspase-3.</t> Cell nuclei were counter-stained by DAPI (blue). Scale bar: 50 µm. Data are representative of 3 independent experiments.
    Cleaved Caspase 3, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 968 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc cleaved caspase 3 cl caspase3 asp175
    Representative costainings of Akirin-2 (green) with a-f. cleaved (c)Caspase-3, g-l. cleaved (c)Caspase-7, and (m-r) cleaved (c)poly(ADP-ribose) polymerase-1 (cPARP-1) (all red) in unexposed, dimethylsulfoxide (DMSO) or TMZ (400 μg/ml, 24 h) treated mock or siAkirin-2 transfected T98G GBM cells Although no clear quantitative data could be obtained, in relation to mock transfected samples (a-c, g-I, m-o) Akirin-2 knock down (d-f, j-l, p-r) resulted in higher amounts of <t>cCaspase-3,</t> -7 or cPARP-1 positively stained cells especially in DMSO and TMZ treated samples. Irrespective of treatment, Akirin-2 knock down became visible by lower intensities and amounts of Akirin-2 positively stained cells. Magnification 400x, bar: 20 μm, representative examples of two independent experiments are shown. For individual images per dye/marker see Supplementary Figs. S5 , S6 and S7 .
    Cleaved Caspase 3 Cl Caspase3 Asp175, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Cell Signaling Technology Inc cleaved caspase3
    Effect of Tristetraprolin (TTP) on cell apoptosis markers (Bax, Bcl-2 and Caspase-3) in HK-2 and NRK-52E cells after treatment of DOX. Western blot for the detection of Bax, Bcl-2, <t>Cleaved-caspase3,</t> Caspase3 and GAPDH in (A) HK-2 and (B) NRK-52E cells. The bar graphs represented the relative expressions compared to GAPDH. All values were expressed as means ± SD, n=5, **P
    Cleaved Caspase3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1999 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc anti cleaved caspase3
    Analysis of GCP proliferation and opoptosis in the LKB1 Atoh1 CKO mice. ( A ) Sections of P3.5 wild-type and LKB1 Atoh1 CKO cerebellum were stained with the anti-PH3 antibody. More PH3 + cells were observed in the mutant cerebellum compared to the wild-type cerebellum, indicating increased GCP proliferation. Red, PH3; Blue, DAPI. Scale bars: 100 μm. ( A’ ) The number of the PH3 + cells per unit size of sagittal sections from the entire cerebellum was quantified in ImageJ (P = 1 × 10 −4 ). ( B ) Immunofluorescence of the BrdU pulse (2 hours) in frozen sections of the P3.5 cerebellum. The cerebellar sections were stained with an anti-BrdU antibody, and more BrdU + cells were observed in the mutant cerebellum compared to the wild-type cerebellum, revealing increased GCP proliferation. Green, BrdU; Blue, DAPI. Scale bars: 100 μm. ( B’ ) The number of BrdU + cells per unit size of sagittal sections from the entire cerebellum was quantified in ImageJ (P = 3 × 10 −4 ). ( C ) Sections of P3.5 wild-type and LKB1 Atoh1 CKO cerebellum were stained with <t>anti-caspase3</t> antibody. No significant change in the number of caspase3 + cells was observed in the mutant cerebellum compared with the wild-type cerebellum. Green, caspase3; Blue, DAPI. Scale bars: 100 μm. ( C’ ) The number of the caspase3 + cells per unit size of sagittal sections from the entire cerebellum was quantified in ImageJ. No significant change in the number of caspase3 + cells were found in the mutant cerebellum compared with the wild-type cerebellum (P = 0.53). The error bars indicate the SEM. *P
    Anti Cleaved Caspase3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 595 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc cleaved caspase3 asp175
    cIAP2 interaction with cleaved <t>caspase-3</t> regulates the conversion of p19 into p17 caspase-3 subunit, its enzymatic activity and apoptosis in microglia. ( a ) Quantification of in situ PLA demonstrating protein interactions between cleaved caspase-3 <t>Asp175</t> and cIAP2 occurring in LPS-treated BV2 microglia cells as compared with untreated cells. Data are presented as mean±S.E.M. of fluorescent dots/cell; n =3; *** P
    Cleaved Caspase3 Asp175, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 139 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Biocare Medical cleaved caspase 3
    Immunohistochemical analyses of MPNST tumors treated with control or DZNep regeant. Represented images of H E staining and immunohistochemical analyses of cell apoptosis marker cleaved <t>caspase</t> 3 and cell proliferation marker Ki-67 in MPNST724 xenograft tumor samples treated with vehicle only, 1 mg/kg DZNep, or 5 mg/kg DZNep. (×200 magnification).
    Cleaved Caspase 3, supplied by Biocare Medical, used in various techniques. Bioz Stars score: 94/100, based on 386 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Santa Cruz Biotechnology cleaved caspase 3
    Immunochemical image for confirmation of reduced cleaved <t>caspase-3</t> expression by DHA treatment. (A) Immunochemical images showed that cleaved caspase-3 positive cells (red) were densely expressed in the EC group. In DHA treatment group, cleaved caspase-3 expression was decreased in rat cortex, compared with the EC group. (B) In DHA treatment group, cleaved caspase-3 positive cells were decreased in rat striatum due to DHA treatment. (C) The graph showed the percentage (%) of cleaved caspase-3 positive cells to compare the difference of cleaved caspase-3 fluorescence intensity. Statistical significance with EC group was determined by t-test. (D) The graph of cleaved caspase-3 protein level showed the same pattern with immunochemical images. Differences were considered significant at * p
    Cleaved Caspase 3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1672 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam cleaved caspase 3
    COQ6 knockdown in cultured podocytes induces apoptosis. (a) Western blot for cleaved <t>caspase-3</t> in the si-NC and si-COQ6 groups. We found that the transfection of siRNA-COQ6 significantly decreased cleaved caspase-3 compared to the negative control. (b) Relative quantification of cleaved caspase-3 (a), normalized to β-actin (* P
    Cleaved Caspase 3, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 2633 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson cleaved caspase 3
    α9β1 inhibits the α3β1-dependent secretion of keratinocyte factors that suppress endothelial cell apoptosis. (A and B) HUVEC apoptosis was measured in response to conditioned media from MK cells that express α3β1 and/or α9β1 in various combinations as indicated. (A, top) Apoptotic cells were detected by immunostaining with an antibody against cleaved <t>caspase</t> 3 (CC3). Bottom, DAPI staining of nuclei. Bar, 100 µm. (B) Graph showing relative caspase 3 activity in HUVECs (EnzChek assay) normalized to the daily mean to account for variability by day. Means ± SEM are shown. n = 3 independent experiments. A one-way analysis of variance with Newman-Keuls’s multiple comparisons test was used. *, P
    Cleaved Caspase 3, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 593 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Proteintech cleaved caspase 3
    CPS-induced apoptosis in ALI cultures of sheep bronchial epithelial cells. Cells were pretreated with/without NAC (10 mM) for 2 h, followed by exposure to indicated conditions for 48 h. (a) 4-week-old ALI cultures of sheep bronchial epithelial cells were apically infected with CPS at 100 ng/ml and MO at MOI of 30 for 48 h before samples were harvested for analysis. Percentages of apoptotic cells were determined by flow cytometry using Annexin V/PI double-staining assay. (b) Immunoblots of apoptosis-associated proteins. The blots were probed for β -actin as a loading control. (c and d) Representative blots for <t>cleaved-caspase-3</t> and cleaved-PARP1 were semiquantified by a densitometric analysis by calculating the fold of change of a protein of interest over β -actin. (e) Relative caspase-3 activity of ALI cells was detected after treating with indicated conditions. Values are mean ± SD for at least three independent experiments performed in triplicate. Compared to non-CPS or MO treatment, ∗ p
    Cleaved Caspase 3, supplied by Proteintech, used in various techniques. Bioz Stars score: 92/100, based on 194 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Covance cleaved caspase 3
    CPS-induced apoptosis in ALI cultures of sheep bronchial epithelial cells. Cells were pretreated with/without NAC (10 mM) for 2 h, followed by exposure to indicated conditions for 48 h. (a) 4-week-old ALI cultures of sheep bronchial epithelial cells were apically infected with CPS at 100 ng/ml and MO at MOI of 30 for 48 h before samples were harvested for analysis. Percentages of apoptotic cells were determined by flow cytometry using Annexin V/PI double-staining assay. (b) Immunoblots of apoptosis-associated proteins. The blots were probed for β -actin as a loading control. (c and d) Representative blots for <t>cleaved-caspase-3</t> and cleaved-PARP1 were semiquantified by a densitometric analysis by calculating the fold of change of a protein of interest over β -actin. (e) Relative caspase-3 activity of ALI cells was detected after treating with indicated conditions. Values are mean ± SD for at least three independent experiments performed in triplicate. Compared to non-CPS or MO treatment, ∗ p
    Cleaved Caspase 3, supplied by Covance, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Beyotime cleaved caspase 3
    Tet and CNTF inhibited the expression of cleaved <t>caspase-3</t> in the retinas 1 day after I/R insult. Notes: Effect of Tet ( A – C , J – L ), CNTF ( D – F , M – O ) and PBS control ( G – I , P – R ) on the expression of cleaved caspase-3 1 day after retinal ischemia. Cleaved caspase-3 is green and DAPI-labeled nuclei are blue in the merged images. Red arrows indicate green fluorescence. The ganglion cell layer, inner nuclear layer, and outer nuclear layer are indicated. Scale bar =20 μm ( A – R ). All images were captured at 40× magnification. Abbreviations: CNTF, ciliary neurotrophic factor; DAPI, 4, 6-diamidino-2-phenylindole; I/R, ischemia/reperfusion; PBS, phosphate-buffered saline; Tet, tetrandrine; GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer.
    Cleaved Caspase 3, supplied by Beyotime, used in various techniques. Bioz Stars score: 93/100, based on 171 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Epitomics cleaved caspase 3
    Effect of TSA on docetaxel‐sensitive and docetaxel‐resistant PC a cells. ( A ) The morphological changes after 24‐hrs treatment with 0, 0.1, 0.2, 0.4 and 0.8 μM of TSA in PC 3 and PC 3/Doc cells. ( B ) Cell viability was determined by the MTT assay after treatment with different TSA concentrations for 24 hrs in PC 3 and PC 3/Doc cells. ( C ) Cell proliferation was examined by EdU incorporation assay at 16 hrs after TSA treatment, and ( D ) EdU‐positive cells were calculated. ( E ) Detection of apoptotic cells after annexin V/ PI staining by flow cytometric analysis after treatment with 0, 0.1, 0.2 and 0.4 μM of TSA for 24 hrs in PC 3/Doc cells and ( F ) the population of the apoptotic cell was calculated. ( G ) Cleaved <t>caspase‐3</t> and PARP expression were examined for cells treated with TSA at various concentrations for 24 hrs. ( H ) Western blot analysis of the cleaved caspase‐3, PARP , rH 2 AX , Rad 51 and Ku70/80 in PC 3/Doc cells treated with TSA (0.4 μM) at different time‐points. ( I ) Morphological changes and ( J ) cell viability in the absence or presence of pan‐caspase inhibitor (z‐ VAD ‐fmk). PC 3/Doc cells were exposed to 10 μM z‐ VAD ‐fmk for 2 hrs prior to TSA (0.4 μM) or vehicle treatment for 24 hrs. * P
    Cleaved Caspase 3, supplied by Epitomics, used in various techniques. Bioz Stars score: 93/100, based on 46 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Cell Signaling Technology Inc rabbit polyclonal anti cleaved caspase3
    Effect of TSA on docetaxel‐sensitive and docetaxel‐resistant PC a cells. ( A ) The morphological changes after 24‐hrs treatment with 0, 0.1, 0.2, 0.4 and 0.8 μM of TSA in PC 3 and PC 3/Doc cells. ( B ) Cell viability was determined by the MTT assay after treatment with different TSA concentrations for 24 hrs in PC 3 and PC 3/Doc cells. ( C ) Cell proliferation was examined by EdU incorporation assay at 16 hrs after TSA treatment, and ( D ) EdU‐positive cells were calculated. ( E ) Detection of apoptotic cells after annexin V/ PI staining by flow cytometric analysis after treatment with 0, 0.1, 0.2 and 0.4 μM of TSA for 24 hrs in PC 3/Doc cells and ( F ) the population of the apoptotic cell was calculated. ( G ) Cleaved <t>caspase‐3</t> and PARP expression were examined for cells treated with TSA at various concentrations for 24 hrs. ( H ) Western blot analysis of the cleaved caspase‐3, PARP , rH 2 AX , Rad 51 and Ku70/80 in PC 3/Doc cells treated with TSA (0.4 μM) at different time‐points. ( I ) Morphological changes and ( J ) cell viability in the absence or presence of pan‐caspase inhibitor (z‐ VAD ‐fmk). PC 3/Doc cells were exposed to 10 μM z‐ VAD ‐fmk for 2 hrs prior to TSA (0.4 μM) or vehicle treatment for 24 hrs. * P
    Rabbit Polyclonal Anti Cleaved Caspase3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 88/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    R&D Systems cleaved caspase 3
    The effect of a DIO regimen +/− EPA+DHA supplementation on IHC tumoral staining. A, representative photomicrographs of pathology and IHC staining of tumors for cleaved <t>caspase-3,</t> COX-2 and phospho-p65. B, bar graphs representing the Aperio image quantitation, scale bars indicate 100 μm, n=6 per group. Means ± SEM, statistically significant (P
    Cleaved Caspase 3, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 393 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Generation of 3D-spheroid GMSCs. ( A ) 4 × 10 4 of GMSCs/well in 200 µl of complete stemgro culture medium was seeded into each well of ultra-low attachment round-shaped 96-U well plates and cultured for 48 h. H E staining of cryosections of GMSC spheroids. Scale bar: 50 µm. ( B ) Immunocytochemistry showed the expression of a panel of MSC-associated markers CD29, CD73 and CD90 and extracellular components such as type I collagen (col-I), vimentin, fibronectin, and laminin in GMSC-derived spheroids. ( C ) GMSC spheroids were dissociated into single cells and stained with Annexin V-FITC and 7-AAD to detect early apoptosis and late apoptotic/necrotic cells by flow cytometric analysis. ( D ) Immunocytochemistry showed that less than 10% of cells inside GMSC spheroids were positive for the cleaved caspase-3. Cell nuclei were counter-stained by DAPI (blue). Scale bar: 50 µm. Data are representative of 3 independent experiments.

    Journal: Scientific Reports

    Article Title: 3D bio-printed scaffold-free nerve constructs with human gingiva-derived mesenchymal stem cells promote rat facial nerve regeneration

    doi: 10.1038/s41598-018-24888-w

    Figure Lengend Snippet: Generation of 3D-spheroid GMSCs. ( A ) 4 × 10 4 of GMSCs/well in 200 µl of complete stemgro culture medium was seeded into each well of ultra-low attachment round-shaped 96-U well plates and cultured for 48 h. H E staining of cryosections of GMSC spheroids. Scale bar: 50 µm. ( B ) Immunocytochemistry showed the expression of a panel of MSC-associated markers CD29, CD73 and CD90 and extracellular components such as type I collagen (col-I), vimentin, fibronectin, and laminin in GMSC-derived spheroids. ( C ) GMSC spheroids were dissociated into single cells and stained with Annexin V-FITC and 7-AAD to detect early apoptosis and late apoptotic/necrotic cells by flow cytometric analysis. ( D ) Immunocytochemistry showed that less than 10% of cells inside GMSC spheroids were positive for the cleaved caspase-3. Cell nuclei were counter-stained by DAPI (blue). Scale bar: 50 µm. Data are representative of 3 independent experiments.

    Article Snippet: Cultured cells fixed with 4% paraformaldehyde (PFA) or cryosections of GMSC spheroids were blocked and permeabilized for 1 h at room temperature in PBS with 2.5% goat serum and 0.5%Triton X‐100, followed by incubation with the following primary antibodies at the appropriate dilution overnight at 4 °C: Nestin (mouse IgG, 1:250) (EMD Millipore, Burlington, MA, USA), CD29 (mouse IgG, 1:250) (BD Bioscience, San Jose, CA, USA), cleaved caspase-3 (Rabbit IgG, 1:250) (EMD Millipore), type I collagen (rabbit IgG, 1:250) (Rockland Biotech, Limerick, PA, USA), CD73(mouse IgG, 1:250) (BD Bioscience), CD90 (mouse IgG, 1:250) (BD Bioscience), vimentin (rabbit IgG, 1:250) (Boster Biological Tech., Pleasanton, CA, USA), fibronectin (rabbit IgG, 1:200 (Sigma, St. Louis, MO, USA), laminin 1 (rabbit IgG, 1:200) (EMD Millipore), β-tubulin III (mouse IgG, 1:200) (BioRad, Hercules, CA, USA), and S-100β (rabbit IgG, 1:250) (Boster Biological Tech).

    Techniques: Cell Culture, Staining, Immunocytochemistry, Expressing, Derivative Assay, Flow Cytometry

    Representative costainings of Akirin-2 (green) with a-f. cleaved (c)Caspase-3, g-l. cleaved (c)Caspase-7, and (m-r) cleaved (c)poly(ADP-ribose) polymerase-1 (cPARP-1) (all red) in unexposed, dimethylsulfoxide (DMSO) or TMZ (400 μg/ml, 24 h) treated mock or siAkirin-2 transfected T98G GBM cells Although no clear quantitative data could be obtained, in relation to mock transfected samples (a-c, g-I, m-o) Akirin-2 knock down (d-f, j-l, p-r) resulted in higher amounts of cCaspase-3, -7 or cPARP-1 positively stained cells especially in DMSO and TMZ treated samples. Irrespective of treatment, Akirin-2 knock down became visible by lower intensities and amounts of Akirin-2 positively stained cells. Magnification 400x, bar: 20 μm, representative examples of two independent experiments are shown. For individual images per dye/marker see Supplementary Figs. S5 , S6 and S7 .

    Journal: Oncotarget

    Article Title: Down regulation of Akirin-2 increases chemosensitivity in human glioblastomas more efficiently than Twist-1

    doi:

    Figure Lengend Snippet: Representative costainings of Akirin-2 (green) with a-f. cleaved (c)Caspase-3, g-l. cleaved (c)Caspase-7, and (m-r) cleaved (c)poly(ADP-ribose) polymerase-1 (cPARP-1) (all red) in unexposed, dimethylsulfoxide (DMSO) or TMZ (400 μg/ml, 24 h) treated mock or siAkirin-2 transfected T98G GBM cells Although no clear quantitative data could be obtained, in relation to mock transfected samples (a-c, g-I, m-o) Akirin-2 knock down (d-f, j-l, p-r) resulted in higher amounts of cCaspase-3, -7 or cPARP-1 positively stained cells especially in DMSO and TMZ treated samples. Irrespective of treatment, Akirin-2 knock down became visible by lower intensities and amounts of Akirin-2 positively stained cells. Magnification 400x, bar: 20 μm, representative examples of two independent experiments are shown. For individual images per dye/marker see Supplementary Figs. S5 , S6 and S7 .

    Article Snippet: The second primary antibody - for transfected/TMZ exposed cells and corresponding controls anti-cPARP (#9541; 1:100; Cell Signaling Technology) or anti-cCaspase-3 (#9661; 1:200, Cell Signaling Technology) or anti-cCaspase-7 (#8438; 1:200, Cell Signaling Technology), for only TMZ exposed cells and corresponding controls vice versa anti-Akirin-2 or anti-Twist-1 - was applied over night at 4°C.

    Techniques: Transfection, Staining, Marker

    ImageStream x Mark II analysis of costainings of Twist-1 (green) with cleaved (c)Caspase-3 (far red) in unexposed, dimethylsulfoxide (DMSO) or TMZ (400 μg/ml, 24 h) treated mock or siTwist-1 transfected T98G GBM cells a-b. A high, medium and a cCaspase-3 negative cell population with variable Twist-1 contents was detectable in all analyzed samples, and c. amounts of high cCaspase-3 positively stained cells increased from unexposed up to DMSO treated samples with higher amounts in siAkirin-2 transfected cells. In TMZ treated samples summarized amounts of high and medium cCaspase-3 positively stained T98 cells were nearly equal including d. a prominent low Twist-1 + medium cCaspase-3 cell population resulting in obliteration of differences between mock and RNAi samples. Representative examples of two independent experiments are shown.

    Journal: Oncotarget

    Article Title: Down regulation of Akirin-2 increases chemosensitivity in human glioblastomas more efficiently than Twist-1

    doi:

    Figure Lengend Snippet: ImageStream x Mark II analysis of costainings of Twist-1 (green) with cleaved (c)Caspase-3 (far red) in unexposed, dimethylsulfoxide (DMSO) or TMZ (400 μg/ml, 24 h) treated mock or siTwist-1 transfected T98G GBM cells a-b. A high, medium and a cCaspase-3 negative cell population with variable Twist-1 contents was detectable in all analyzed samples, and c. amounts of high cCaspase-3 positively stained cells increased from unexposed up to DMSO treated samples with higher amounts in siAkirin-2 transfected cells. In TMZ treated samples summarized amounts of high and medium cCaspase-3 positively stained T98 cells were nearly equal including d. a prominent low Twist-1 + medium cCaspase-3 cell population resulting in obliteration of differences between mock and RNAi samples. Representative examples of two independent experiments are shown.

    Article Snippet: The second primary antibody - for transfected/TMZ exposed cells and corresponding controls anti-cPARP (#9541; 1:100; Cell Signaling Technology) or anti-cCaspase-3 (#9661; 1:200, Cell Signaling Technology) or anti-cCaspase-7 (#8438; 1:200, Cell Signaling Technology), for only TMZ exposed cells and corresponding controls vice versa anti-Akirin-2 or anti-Twist-1 - was applied over night at 4°C.

    Techniques: Transfection, Staining

    ImageStream x Mark II analysis of costainings of Akirin-2 (green) with cleaved (c)Caspase-3 (far red) in unexposed, dimethylsulfoxide (DMSO) or TMZ (400μg/ml, 24 h) treated mock or siAkirin-2 knock down (kd) T98G GBM cells a, b. A high, medium and a cCaspase-3 negative cell population with variable Akirin-2 contents was detectable in all analyzed samples, and c. amounts of high cCaspase-3 positively stained cells increased from unexposed up to TMZ treated samples with higher amounts in siAkirin-2 transfected cells. d. Higher contents of a low Akirin-2 + high cCaspase-3 positive cell population were found for siAkirin-2 transfected cells in unexposed, DMSO and TMZ treated samples. Representative examples of two independent experiments are shown.

    Journal: Oncotarget

    Article Title: Down regulation of Akirin-2 increases chemosensitivity in human glioblastomas more efficiently than Twist-1

    doi:

    Figure Lengend Snippet: ImageStream x Mark II analysis of costainings of Akirin-2 (green) with cleaved (c)Caspase-3 (far red) in unexposed, dimethylsulfoxide (DMSO) or TMZ (400μg/ml, 24 h) treated mock or siAkirin-2 knock down (kd) T98G GBM cells a, b. A high, medium and a cCaspase-3 negative cell population with variable Akirin-2 contents was detectable in all analyzed samples, and c. amounts of high cCaspase-3 positively stained cells increased from unexposed up to TMZ treated samples with higher amounts in siAkirin-2 transfected cells. d. Higher contents of a low Akirin-2 + high cCaspase-3 positive cell population were found for siAkirin-2 transfected cells in unexposed, DMSO and TMZ treated samples. Representative examples of two independent experiments are shown.

    Article Snippet: The second primary antibody - for transfected/TMZ exposed cells and corresponding controls anti-cPARP (#9541; 1:100; Cell Signaling Technology) or anti-cCaspase-3 (#9661; 1:200, Cell Signaling Technology) or anti-cCaspase-7 (#8438; 1:200, Cell Signaling Technology), for only TMZ exposed cells and corresponding controls vice versa anti-Akirin-2 or anti-Twist-1 - was applied over night at 4°C.

    Techniques: Staining, Transfection

    Akirin-2 mediated chemoresistance could be abolished by RNAi technology as measured by (a) cleaved (c)Caspase-3/-7 activity assay, (b) cCaspase-3 and -7 and (c) cleaved (c)poly(ADP-ribose) polymerase-1 (cPARP-1) Western Blots a. In relation to mock transfected T98G cells Akirin-2 knock down (kd) was able to induce cell death to significantly greater extents in dimethylsulfoxide (DMSO) as well as in TMZ (400 μg/ml, 24 h) treated samples (n = 6; triple values; boxplot: bold line = median; box = upper and lower quartile; whisker = 1.5-fold of interquartile range, circles = outlier). b. Results were confirmed by Western Blot using specific antibodies directed against cCaspase-3 and -7, respectively. c. PARP-1 Western Blot using two different antibodies specifically directed against cleaved PARP-1 (bottom) or both uncleaved and cleaved PARP-1 (top) additionally approved results. Equal protein loading was confirmed by detection of glycerinaldehyde-3-phosphate-dehydrogenase (GAPDH) or heat shock protein 90 (Hsp90), and efficiency of knock down was proven by qRT-PCR for all experiments in parallel. Representative examples of two independent experiments are shown.

    Journal: Oncotarget

    Article Title: Down regulation of Akirin-2 increases chemosensitivity in human glioblastomas more efficiently than Twist-1

    doi:

    Figure Lengend Snippet: Akirin-2 mediated chemoresistance could be abolished by RNAi technology as measured by (a) cleaved (c)Caspase-3/-7 activity assay, (b) cCaspase-3 and -7 and (c) cleaved (c)poly(ADP-ribose) polymerase-1 (cPARP-1) Western Blots a. In relation to mock transfected T98G cells Akirin-2 knock down (kd) was able to induce cell death to significantly greater extents in dimethylsulfoxide (DMSO) as well as in TMZ (400 μg/ml, 24 h) treated samples (n = 6; triple values; boxplot: bold line = median; box = upper and lower quartile; whisker = 1.5-fold of interquartile range, circles = outlier). b. Results were confirmed by Western Blot using specific antibodies directed against cCaspase-3 and -7, respectively. c. PARP-1 Western Blot using two different antibodies specifically directed against cleaved PARP-1 (bottom) or both uncleaved and cleaved PARP-1 (top) additionally approved results. Equal protein loading was confirmed by detection of glycerinaldehyde-3-phosphate-dehydrogenase (GAPDH) or heat shock protein 90 (Hsp90), and efficiency of knock down was proven by qRT-PCR for all experiments in parallel. Representative examples of two independent experiments are shown.

    Article Snippet: The second primary antibody - for transfected/TMZ exposed cells and corresponding controls anti-cPARP (#9541; 1:100; Cell Signaling Technology) or anti-cCaspase-3 (#9661; 1:200, Cell Signaling Technology) or anti-cCaspase-7 (#8438; 1:200, Cell Signaling Technology), for only TMZ exposed cells and corresponding controls vice versa anti-Akirin-2 or anti-Twist-1 - was applied over night at 4°C.

    Techniques: Activity Assay, Western Blot, Transfection, Whisker Assay, Quantitative RT-PCR

    Representative costainings of Twist-1 (green) with a-f. cleaved (c)Caspase-3, g-l. cleaved (c)Caspase-7, and m-r. cleaved (c)poly(ADP-ribose) polymerase-1 (cPARP-1) (all red) in unexposed, dimethylsulfoxide (DMSO) or TMZ (400 μg/ml, 24 h) treated mock or siTwist-1 transfected T98G GBM cells. Although clear quantitative data could not be obtained, in relation to mock transfected samples (a-b, g-h, m-n) Twist-1 knock down (d-e, j-k, p-q) resulted in higher amounts of cCaspase-3, -7 or cPARP-1 positively stained cells in unexposed and DMSO treated samples, respectively, whereas in TMZ treated ones (c vs. f, i vs. l, o vs. r) no distinct differences were detectable Twist-1 knock down became visible by lower intensities and amounts of Twist-1 positively stained cells in unexposed and DMSO treated samples, with TMZ application Twist-1 knock down seemed to be antagonized. Magnification 400x, bar: 20 μm, representative examples of two independent experiments are shown. For individual images per dye/marker see Supplementary Figs. S8 , S9 and S10 .

    Journal: Oncotarget

    Article Title: Down regulation of Akirin-2 increases chemosensitivity in human glioblastomas more efficiently than Twist-1

    doi:

    Figure Lengend Snippet: Representative costainings of Twist-1 (green) with a-f. cleaved (c)Caspase-3, g-l. cleaved (c)Caspase-7, and m-r. cleaved (c)poly(ADP-ribose) polymerase-1 (cPARP-1) (all red) in unexposed, dimethylsulfoxide (DMSO) or TMZ (400 μg/ml, 24 h) treated mock or siTwist-1 transfected T98G GBM cells. Although clear quantitative data could not be obtained, in relation to mock transfected samples (a-b, g-h, m-n) Twist-1 knock down (d-e, j-k, p-q) resulted in higher amounts of cCaspase-3, -7 or cPARP-1 positively stained cells in unexposed and DMSO treated samples, respectively, whereas in TMZ treated ones (c vs. f, i vs. l, o vs. r) no distinct differences were detectable Twist-1 knock down became visible by lower intensities and amounts of Twist-1 positively stained cells in unexposed and DMSO treated samples, with TMZ application Twist-1 knock down seemed to be antagonized. Magnification 400x, bar: 20 μm, representative examples of two independent experiments are shown. For individual images per dye/marker see Supplementary Figs. S8 , S9 and S10 .

    Article Snippet: The second primary antibody - for transfected/TMZ exposed cells and corresponding controls anti-cPARP (#9541; 1:100; Cell Signaling Technology) or anti-cCaspase-3 (#9661; 1:200, Cell Signaling Technology) or anti-cCaspase-7 (#8438; 1:200, Cell Signaling Technology), for only TMZ exposed cells and corresponding controls vice versa anti-Akirin-2 or anti-Twist-1 - was applied over night at 4°C.

    Techniques: Transfection, Staining, Marker

    Effect of Tristetraprolin (TTP) on cell apoptosis markers (Bax, Bcl-2 and Caspase-3) in HK-2 and NRK-52E cells after treatment of DOX. Western blot for the detection of Bax, Bcl-2, Cleaved-caspase3, Caspase3 and GAPDH in (A) HK-2 and (B) NRK-52E cells. The bar graphs represented the relative expressions compared to GAPDH. All values were expressed as means ± SD, n=5, **P

    Journal: American Journal of Translational Research

    Article Title: Effects of tristetraprolin on doxorubicin (adriamycin)-induced experimental kidney injury through inhibiting IL-13/STAT6 signal pathway

    doi:

    Figure Lengend Snippet: Effect of Tristetraprolin (TTP) on cell apoptosis markers (Bax, Bcl-2 and Caspase-3) in HK-2 and NRK-52E cells after treatment of DOX. Western blot for the detection of Bax, Bcl-2, Cleaved-caspase3, Caspase3 and GAPDH in (A) HK-2 and (B) NRK-52E cells. The bar graphs represented the relative expressions compared to GAPDH. All values were expressed as means ± SD, n=5, **P

    Article Snippet: Thereafter, the PVDF membranes were hybridized in blocking buffer overnight at 4°C together with primary antibodies, including TTP (71632; 1:1000; Cell Signaling Technology, Inc.), Kim-1 (ab47635; 1:1000; Abcam), TNF-α (3707; 1:1000; Cell Signaling Technology, Inc.), IL-1β (12242; 1:1000; Cell Signaling Technology, Inc.), IL-6 (12912; 1:1000; Cell Signaling Technology, Inc.), IL-13 (ab106732; 1:1000; Abcam), p-STAT6 (56554; 1:1000; Cell Signaling Technology, Inc.), STAT6 (5397; 1:1000; Cell Signaling Technology, Inc.), cleaved-caspase3 (9661; 1:1000; Cell Signaling Technology, Inc.), caspase3 (9662; 1:1000; Cell Signaling Technology, Inc.), Bax (ab32503; 1:1,000; Abcam) and Bcl-2 (ab196495; 1:1,000; Abcam).

    Techniques: Western Blot

    Combined ARS1620/SHP2 inhibition is highly efficacious in PDAC models in vivo . A , Pancreas tumors were established in syngeneic mice by orthotopic injections of KCP cells, and 14 days later, mice were treated with vehicle, SHP099, ARS1620 or both drugs (Combo), as depicted. Tumor weight was quantified in a cohort at Day 0 (baseline) and in treated mice at Day 10. B , Immunoblots of KCP-derived tumor lysates showing effects of the indicated treatments on KRAS G12C -GTP, pERK, and DUSP6 levels. C , ERK-dependent gene expression, assessed by RNAseq, in KCP tumors treated for 3 days, as indicated in A (colors indicate log2FC). D-E , Time-dependent increase in RTK (D) and RTK ligands (E) gene expression in KCP-derived orthotopic tumors after vehicle, SHP099, ARS1620 and Combo treatment at Day 3, determined by RNAseq (colors represent log2FC). F , H E, Masson Trichome, CD31, pERK, Ki67 and cleaved Caspase 3 staining and quantification in KCP tumor sections from mice after 10 days of treatment, as indicated. G , KCP tumors were established in syngeneic mice and allowed to grow to much larger size before treatments were initiated, as depicted in the scheme. Tumor weight was quantified in one cohort before treatment, in another cohort after 12 days of treatment, and after drug withdrawal, at Day 27, as indicated. H , Kaplan-Meier curve of KCP tumor-bearing mice after withdrawal of the indicated drugs (top). Tumor growth curve after withdrawal of indicated treatment at day 12 (bottom). H , Response of sub-cutaneous NY53 patient-derived xenograft to treatment with vehicle, SHP099, ARS1620 or both drugs. For all experiments, drug doses were: SHP099 (75 mg/kg body weight, daily), ARS1620 (200 mg/kg body weight, daily) or both drugs (daily). Data represent mean ± SD; *P

    Journal: bioRxiv

    Article Title: SHP2 Inhibition Abrogates Adaptive Resistance to KRASG12C-Inhibition and Remodels the Tumor Microenvironment of KRAS-Mutant Tumors

    doi: 10.1101/2020.05.30.125138

    Figure Lengend Snippet: Combined ARS1620/SHP2 inhibition is highly efficacious in PDAC models in vivo . A , Pancreas tumors were established in syngeneic mice by orthotopic injections of KCP cells, and 14 days later, mice were treated with vehicle, SHP099, ARS1620 or both drugs (Combo), as depicted. Tumor weight was quantified in a cohort at Day 0 (baseline) and in treated mice at Day 10. B , Immunoblots of KCP-derived tumor lysates showing effects of the indicated treatments on KRAS G12C -GTP, pERK, and DUSP6 levels. C , ERK-dependent gene expression, assessed by RNAseq, in KCP tumors treated for 3 days, as indicated in A (colors indicate log2FC). D-E , Time-dependent increase in RTK (D) and RTK ligands (E) gene expression in KCP-derived orthotopic tumors after vehicle, SHP099, ARS1620 and Combo treatment at Day 3, determined by RNAseq (colors represent log2FC). F , H E, Masson Trichome, CD31, pERK, Ki67 and cleaved Caspase 3 staining and quantification in KCP tumor sections from mice after 10 days of treatment, as indicated. G , KCP tumors were established in syngeneic mice and allowed to grow to much larger size before treatments were initiated, as depicted in the scheme. Tumor weight was quantified in one cohort before treatment, in another cohort after 12 days of treatment, and after drug withdrawal, at Day 27, as indicated. H , Kaplan-Meier curve of KCP tumor-bearing mice after withdrawal of the indicated drugs (top). Tumor growth curve after withdrawal of indicated treatment at day 12 (bottom). H , Response of sub-cutaneous NY53 patient-derived xenograft to treatment with vehicle, SHP099, ARS1620 or both drugs. For all experiments, drug doses were: SHP099 (75 mg/kg body weight, daily), ARS1620 (200 mg/kg body weight, daily) or both drugs (daily). Data represent mean ± SD; *P

    Article Snippet: IHC for pERK (Cell Signaling, 4370), CD31 (Cell Signaling, D8V9E), Cleaved Caspase 3 (Cell Signaling, D3E9), Ki67 (Spring Biosciences, SP6), αSMA (Abcam, ab5694) was performed on sections from paraformaldehyde-fixed tumors.

    Techniques: Inhibition, In Vivo, Mouse Assay, Western Blot, Derivative Assay, Expressing, Staining

    Analysis of GCP proliferation and opoptosis in the LKB1 Atoh1 CKO mice. ( A ) Sections of P3.5 wild-type and LKB1 Atoh1 CKO cerebellum were stained with the anti-PH3 antibody. More PH3 + cells were observed in the mutant cerebellum compared to the wild-type cerebellum, indicating increased GCP proliferation. Red, PH3; Blue, DAPI. Scale bars: 100 μm. ( A’ ) The number of the PH3 + cells per unit size of sagittal sections from the entire cerebellum was quantified in ImageJ (P = 1 × 10 −4 ). ( B ) Immunofluorescence of the BrdU pulse (2 hours) in frozen sections of the P3.5 cerebellum. The cerebellar sections were stained with an anti-BrdU antibody, and more BrdU + cells were observed in the mutant cerebellum compared to the wild-type cerebellum, revealing increased GCP proliferation. Green, BrdU; Blue, DAPI. Scale bars: 100 μm. ( B’ ) The number of BrdU + cells per unit size of sagittal sections from the entire cerebellum was quantified in ImageJ (P = 3 × 10 −4 ). ( C ) Sections of P3.5 wild-type and LKB1 Atoh1 CKO cerebellum were stained with anti-caspase3 antibody. No significant change in the number of caspase3 + cells was observed in the mutant cerebellum compared with the wild-type cerebellum. Green, caspase3; Blue, DAPI. Scale bars: 100 μm. ( C’ ) The number of the caspase3 + cells per unit size of sagittal sections from the entire cerebellum was quantified in ImageJ. No significant change in the number of caspase3 + cells were found in the mutant cerebellum compared with the wild-type cerebellum (P = 0.53). The error bars indicate the SEM. *P

    Journal: Scientific Reports

    Article Title: LKB1 Regulates Cerebellar Development by Controlling Sonic Hedgehog-mediated Granule Cell Precursor Proliferation and Granule Cell Migration

    doi: 10.1038/srep16232

    Figure Lengend Snippet: Analysis of GCP proliferation and opoptosis in the LKB1 Atoh1 CKO mice. ( A ) Sections of P3.5 wild-type and LKB1 Atoh1 CKO cerebellum were stained with the anti-PH3 antibody. More PH3 + cells were observed in the mutant cerebellum compared to the wild-type cerebellum, indicating increased GCP proliferation. Red, PH3; Blue, DAPI. Scale bars: 100 μm. ( A’ ) The number of the PH3 + cells per unit size of sagittal sections from the entire cerebellum was quantified in ImageJ (P = 1 × 10 −4 ). ( B ) Immunofluorescence of the BrdU pulse (2 hours) in frozen sections of the P3.5 cerebellum. The cerebellar sections were stained with an anti-BrdU antibody, and more BrdU + cells were observed in the mutant cerebellum compared to the wild-type cerebellum, revealing increased GCP proliferation. Green, BrdU; Blue, DAPI. Scale bars: 100 μm. ( B’ ) The number of BrdU + cells per unit size of sagittal sections from the entire cerebellum was quantified in ImageJ (P = 3 × 10 −4 ). ( C ) Sections of P3.5 wild-type and LKB1 Atoh1 CKO cerebellum were stained with anti-caspase3 antibody. No significant change in the number of caspase3 + cells was observed in the mutant cerebellum compared with the wild-type cerebellum. Green, caspase3; Blue, DAPI. Scale bars: 100 μm. ( C’ ) The number of the caspase3 + cells per unit size of sagittal sections from the entire cerebellum was quantified in ImageJ. No significant change in the number of caspase3 + cells were found in the mutant cerebellum compared with the wild-type cerebellum (P = 0.53). The error bars indicate the SEM. *P

    Article Snippet: Primary antibodies included anti-LKB1 (1:200, Upstate), anti-BrdU (1:400, Sigma), anti-PH3 (1:400, Bioworlde), anti-NeuN (1:200, Abcam), anti-NeuN (1:500, Cell Signaling Technology), anti-GFAP (1:200, Cell Signaling Technology), anti-calbindinD28K (1:1000, Abcam), anti-Cleaved Caspase3 (1:400, CST), anti-Tbr2 (1:200, Abcam) and anti-cyclinD1 (1:500, Abcam).

    Techniques: Mouse Assay, Staining, Mutagenesis, Immunofluorescence

    cIAP2 interaction with cleaved caspase-3 regulates the conversion of p19 into p17 caspase-3 subunit, its enzymatic activity and apoptosis in microglia. ( a ) Quantification of in situ PLA demonstrating protein interactions between cleaved caspase-3 Asp175 and cIAP2 occurring in LPS-treated BV2 microglia cells as compared with untreated cells. Data are presented as mean±S.E.M. of fluorescent dots/cell; n =3; *** P

    Journal: Cell Death & Disease

    Article Title: Regulation of caspase-3 processing by cIAP2 controls the switch between pro-inflammatory activation and cell death in microglia

    doi: 10.1038/cddis.2014.514

    Figure Lengend Snippet: cIAP2 interaction with cleaved caspase-3 regulates the conversion of p19 into p17 caspase-3 subunit, its enzymatic activity and apoptosis in microglia. ( a ) Quantification of in situ PLA demonstrating protein interactions between cleaved caspase-3 Asp175 and cIAP2 occurring in LPS-treated BV2 microglia cells as compared with untreated cells. Data are presented as mean±S.E.M. of fluorescent dots/cell; n =3; *** P

    Article Snippet: Membranes were blocked in 5% milk and incubated with antibodies raised against cIAP2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), cleaved caspase-3 (Asp175) (Cell Signaling, Beverly, MA, USA), cleaved PARP (Cell Signaling), G3PDH (Trevigen, Gaithersburg, MD, USA), iNOS (Santa Cruz Biotechnology), Lamin B (Abcam, Cambridge, MA, USA) or H4 (Active Motif, Carlsbad, CA, USA), overnight at 4 °C, followed by incubation with the appropriate horseradish peroxidase secondary antibody (Pierce, Rockford, IL, USA, 1 : 10 000) for 1 h at room temperature.

    Techniques: Activity Assay, In Situ, Proximity Ligation Assay

    Scheme illustrating the effect of cIAP2 on the caspase-3 activation steps and consequently biological functions. Pro-caspase-3 is cleaved by upstream caspases, such as active caspases-8, at Asp175 to generate intermediate, yet still active, p19/p12 complexes. Thereafter, autocatalytic processing at residue Asp28 and removal of the short prodomain from the p19 peptides, generate p17/p12 complexes that form the fully mature form of the enzyme and translocate to the nucleus. In turn, p19/p12 and p17/p12 complexes can cleave substrates in the cytoplasmic or cytoplasmic/nuclear cell compartment, respectively, and thereby regulate pro-inflammatory activation or apoptotic cell death of the microglia cells. Upon pro-inflammatory stimulation, upregulated cIAP2 binds to caspase-3 prodomain and prevents the p19 to p17 caspase-3 subunit conversion

    Journal: Cell Death & Disease

    Article Title: Regulation of caspase-3 processing by cIAP2 controls the switch between pro-inflammatory activation and cell death in microglia

    doi: 10.1038/cddis.2014.514

    Figure Lengend Snippet: Scheme illustrating the effect of cIAP2 on the caspase-3 activation steps and consequently biological functions. Pro-caspase-3 is cleaved by upstream caspases, such as active caspases-8, at Asp175 to generate intermediate, yet still active, p19/p12 complexes. Thereafter, autocatalytic processing at residue Asp28 and removal of the short prodomain from the p19 peptides, generate p17/p12 complexes that form the fully mature form of the enzyme and translocate to the nucleus. In turn, p19/p12 and p17/p12 complexes can cleave substrates in the cytoplasmic or cytoplasmic/nuclear cell compartment, respectively, and thereby regulate pro-inflammatory activation or apoptotic cell death of the microglia cells. Upon pro-inflammatory stimulation, upregulated cIAP2 binds to caspase-3 prodomain and prevents the p19 to p17 caspase-3 subunit conversion

    Article Snippet: Membranes were blocked in 5% milk and incubated with antibodies raised against cIAP2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), cleaved caspase-3 (Asp175) (Cell Signaling, Beverly, MA, USA), cleaved PARP (Cell Signaling), G3PDH (Trevigen, Gaithersburg, MD, USA), iNOS (Santa Cruz Biotechnology), Lamin B (Abcam, Cambridge, MA, USA) or H4 (Active Motif, Carlsbad, CA, USA), overnight at 4 °C, followed by incubation with the appropriate horseradish peroxidase secondary antibody (Pierce, Rockford, IL, USA, 1 : 10 000) for 1 h at room temperature.

    Techniques: Activation Assay

    Distinctive caspase-3 processing profile in pro-inflammatory activated versus dying microglia. BV2 microglia cells were treated with 1 μ g/ml LPS for 24 h or 0.1 μ M STS for 3 h to promote pro-inflammatory activation or cell death, respectively. ( a ) Immunoblot analysis demonstrates presence of the p19 caspase-3 subunit in the immune complexes formed after pull down with cleaved caspase-3 Asp175 antibody, which recognizes both p17 and p19 subunits, in LPS-treated microglia. Immune complexes from STS-treated microglia contained both p17 and p19 caspase-3 subunits. ( b ) Subcellular fractionation illustrates the restricted cytoplasmic localization of p19 subunit, in contrast to the p17 subunit that was found to localize in both the cytoplasmic and the nuclear fractions

    Journal: Cell Death & Disease

    Article Title: Regulation of caspase-3 processing by cIAP2 controls the switch between pro-inflammatory activation and cell death in microglia

    doi: 10.1038/cddis.2014.514

    Figure Lengend Snippet: Distinctive caspase-3 processing profile in pro-inflammatory activated versus dying microglia. BV2 microglia cells were treated with 1 μ g/ml LPS for 24 h or 0.1 μ M STS for 3 h to promote pro-inflammatory activation or cell death, respectively. ( a ) Immunoblot analysis demonstrates presence of the p19 caspase-3 subunit in the immune complexes formed after pull down with cleaved caspase-3 Asp175 antibody, which recognizes both p17 and p19 subunits, in LPS-treated microglia. Immune complexes from STS-treated microglia contained both p17 and p19 caspase-3 subunits. ( b ) Subcellular fractionation illustrates the restricted cytoplasmic localization of p19 subunit, in contrast to the p17 subunit that was found to localize in both the cytoplasmic and the nuclear fractions

    Article Snippet: Membranes were blocked in 5% milk and incubated with antibodies raised against cIAP2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), cleaved caspase-3 (Asp175) (Cell Signaling, Beverly, MA, USA), cleaved PARP (Cell Signaling), G3PDH (Trevigen, Gaithersburg, MD, USA), iNOS (Santa Cruz Biotechnology), Lamin B (Abcam, Cambridge, MA, USA) or H4 (Active Motif, Carlsbad, CA, USA), overnight at 4 °C, followed by incubation with the appropriate horseradish peroxidase secondary antibody (Pierce, Rockford, IL, USA, 1 : 10 000) for 1 h at room temperature.

    Techniques: Activation Assay, Fractionation

    SMAC mimetic enhances caspase-3 activation and promotes cell death of LPS-activated microglia. BV2 microglia cells, pretreated or not with 1 μ M BV6 compound for 24 h, were subsequently treated with 1 μ g/ml LPS for 6 h. STS (0.1 μ M) for 3 h was used as cell death stimulus. ( a ) cIAP2 expression was assessed by FACS analysis. Pretreatment with the BV6 SMAC mimetic compound led to ( b ) increased appearance of cleaved caspase-3 Asp175 as seen by FACS analysis and ( c ) caspase-3 activity (DEVD-ase activity), ( d ) decreased IL-1 β mRNA expression and ( e ) cell death as monitored by the appearance of fragmented, damaged or condensed nuclei in LPS-treated BV2 microglia cells. Data are expressed as mean±S.E.M.; n =3; * P

    Journal: Cell Death & Disease

    Article Title: Regulation of caspase-3 processing by cIAP2 controls the switch between pro-inflammatory activation and cell death in microglia

    doi: 10.1038/cddis.2014.514

    Figure Lengend Snippet: SMAC mimetic enhances caspase-3 activation and promotes cell death of LPS-activated microglia. BV2 microglia cells, pretreated or not with 1 μ M BV6 compound for 24 h, were subsequently treated with 1 μ g/ml LPS for 6 h. STS (0.1 μ M) for 3 h was used as cell death stimulus. ( a ) cIAP2 expression was assessed by FACS analysis. Pretreatment with the BV6 SMAC mimetic compound led to ( b ) increased appearance of cleaved caspase-3 Asp175 as seen by FACS analysis and ( c ) caspase-3 activity (DEVD-ase activity), ( d ) decreased IL-1 β mRNA expression and ( e ) cell death as monitored by the appearance of fragmented, damaged or condensed nuclei in LPS-treated BV2 microglia cells. Data are expressed as mean±S.E.M.; n =3; * P

    Article Snippet: Membranes were blocked in 5% milk and incubated with antibodies raised against cIAP2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), cleaved caspase-3 (Asp175) (Cell Signaling, Beverly, MA, USA), cleaved PARP (Cell Signaling), G3PDH (Trevigen, Gaithersburg, MD, USA), iNOS (Santa Cruz Biotechnology), Lamin B (Abcam, Cambridge, MA, USA) or H4 (Active Motif, Carlsbad, CA, USA), overnight at 4 °C, followed by incubation with the appropriate horseradish peroxidase secondary antibody (Pierce, Rockford, IL, USA, 1 : 10 000) for 1 h at room temperature.

    Techniques: Activation Assay, Expressing, FACS, Activity Assay

    Immunohistochemical analyses of MPNST tumors treated with control or DZNep regeant. Represented images of H E staining and immunohistochemical analyses of cell apoptosis marker cleaved caspase 3 and cell proliferation marker Ki-67 in MPNST724 xenograft tumor samples treated with vehicle only, 1 mg/kg DZNep, or 5 mg/kg DZNep. (×200 magnification).

    Journal: Molecular Cancer

    Article Title: Antitumor effects of pharmacological EZH2 inhibition on malignant peripheral nerve sheath tumor through the miR-30a and KPNB1 pathway

    doi: 10.1186/s12943-015-0325-1

    Figure Lengend Snippet: Immunohistochemical analyses of MPNST tumors treated with control or DZNep regeant. Represented images of H E staining and immunohistochemical analyses of cell apoptosis marker cleaved caspase 3 and cell proliferation marker Ki-67 in MPNST724 xenograft tumor samples treated with vehicle only, 1 mg/kg DZNep, or 5 mg/kg DZNep. (×200 magnification).

    Article Snippet: Commercially available antibodies were used for all immunoblot and immunohistochemical detection of EZH2 (1:1000, D2C9, Cell Signaling ) , KPNB1 (1:5000, NB100-81650, Novus Biologicals,), cleaved PARP (1:1000, ab32064, Abcam), vimentin (1:2000, RV202, Santa Cruz), E-cadherin (1:1000, H-108, Santa Cruz), Ki-67 (1:1000, MIB-1; Dako), cleaved caspase 3 (1:1000, BioCare Medical), GAPDH-HRP (1:5000, ab9483, Abcam), and actin-HRP (1:5000, I-19, Santa Cruz).

    Techniques: Immunohistochemistry, Staining, Marker

    Immunochemical image for confirmation of reduced cleaved caspase-3 expression by DHA treatment. (A) Immunochemical images showed that cleaved caspase-3 positive cells (red) were densely expressed in the EC group. In DHA treatment group, cleaved caspase-3 expression was decreased in rat cortex, compared with the EC group. (B) In DHA treatment group, cleaved caspase-3 positive cells were decreased in rat striatum due to DHA treatment. (C) The graph showed the percentage (%) of cleaved caspase-3 positive cells to compare the difference of cleaved caspase-3 fluorescence intensity. Statistical significance with EC group was determined by t-test. (D) The graph of cleaved caspase-3 protein level showed the same pattern with immunochemical images. Differences were considered significant at * p

    Journal: Experimental Neurobiology

    Article Title: Dehydroascorbic Acid Attenuates Ischemic Brain Edema and Neurotoxicity in Cerebral Ischemia: An in vivo Study

    doi: 10.5607/en.2015.24.1.41

    Figure Lengend Snippet: Immunochemical image for confirmation of reduced cleaved caspase-3 expression by DHA treatment. (A) Immunochemical images showed that cleaved caspase-3 positive cells (red) were densely expressed in the EC group. In DHA treatment group, cleaved caspase-3 expression was decreased in rat cortex, compared with the EC group. (B) In DHA treatment group, cleaved caspase-3 positive cells were decreased in rat striatum due to DHA treatment. (C) The graph showed the percentage (%) of cleaved caspase-3 positive cells to compare the difference of cleaved caspase-3 fluorescence intensity. Statistical significance with EC group was determined by t-test. (D) The graph of cleaved caspase-3 protein level showed the same pattern with immunochemical images. Differences were considered significant at * p

    Article Snippet: Taken together, our findings suggest three points that 1) DHA is involved in the inhibition of AQP-1 expression and the preservation of claudin 5, ultimately resulting in the reduction of edema formation induced by cerebral ischemia, 2) DHA is associated with the decrease of Bax, cleaved caspase-3 and iNOS expression, ultimately resulting in the protection of cell death against neurotoxicity following cerebral ischemia, 3) DHA is linked to the preservation of PSD-95 protein expression, ultimately resulting in the improvement of neuron's synaptic connection in cerebral ischemia.

    Techniques: Expressing, Fluorescence

    COQ6 knockdown in cultured podocytes induces apoptosis. (a) Western blot for cleaved caspase-3 in the si-NC and si-COQ6 groups. We found that the transfection of siRNA-COQ6 significantly decreased cleaved caspase-3 compared to the negative control. (b) Relative quantification of cleaved caspase-3 (a), normalized to β-actin (* P

    Journal: Chinese Medical Journal

    Article Title: New Mutation of Coenzyme Q10 Monooxygenase 6 Causing Podocyte Injury in a Focal Segmental Glomerulosclerosis Patient

    doi: 10.4103/0366-6999.245158

    Figure Lengend Snippet: COQ6 knockdown in cultured podocytes induces apoptosis. (a) Western blot for cleaved caspase-3 in the si-NC and si-COQ6 groups. We found that the transfection of siRNA-COQ6 significantly decreased cleaved caspase-3 compared to the negative control. (b) Relative quantification of cleaved caspase-3 (a), normalized to β-actin (* P

    Article Snippet: An increase in cleaved caspase-3 was observed in the si-COQ6 group compared to the control group [Figure and ].

    Techniques: Cell Culture, Western Blot, Transfection, Negative Control

    α9β1 inhibits the α3β1-dependent secretion of keratinocyte factors that suppress endothelial cell apoptosis. (A and B) HUVEC apoptosis was measured in response to conditioned media from MK cells that express α3β1 and/or α9β1 in various combinations as indicated. (A, top) Apoptotic cells were detected by immunostaining with an antibody against cleaved caspase 3 (CC3). Bottom, DAPI staining of nuclei. Bar, 100 µm. (B) Graph showing relative caspase 3 activity in HUVECs (EnzChek assay) normalized to the daily mean to account for variability by day. Means ± SEM are shown. n = 3 independent experiments. A one-way analysis of variance with Newman-Keuls’s multiple comparisons test was used. *, P

    Journal: The Journal of Cell Biology

    Article Title: Suppression of integrin α3β1 by α9β1 in the epidermis controls the paracrine resolution of wound angiogenesis

    doi: 10.1083/jcb.201510042

    Figure Lengend Snippet: α9β1 inhibits the α3β1-dependent secretion of keratinocyte factors that suppress endothelial cell apoptosis. (A and B) HUVEC apoptosis was measured in response to conditioned media from MK cells that express α3β1 and/or α9β1 in various combinations as indicated. (A, top) Apoptotic cells were detected by immunostaining with an antibody against cleaved caspase 3 (CC3). Bottom, DAPI staining of nuclei. Bar, 100 µm. (B) Graph showing relative caspase 3 activity in HUVECs (EnzChek assay) normalized to the daily mean to account for variability by day. Means ± SEM are shown. n = 3 independent experiments. A one-way analysis of variance with Newman-Keuls’s multiple comparisons test was used. *, P

    Article Snippet: Immunohistology of tissue sections 10-µm frozen tissue sections were rehydrated in 0.2% Tween 20/PBS for 10 min, fixed in 3.7% formaldehyde, permeabilized in 0.5% Triton X-100, blocked in 10% heat-inactivated goat serum/5% milk/PBS for 1 h, and then stained with anti–cleaved caspase 3, anti-CD31 (BD), anti–LN-332 (Abcam), anti-FN (Sigma-Aldrich), anti–MMP-9 (Sigma-Aldrich), or anti–keratin 14 (Covance).

    Techniques: Immunostaining, Staining, Activity Assay

    Persistent vasculature in wounds of α9eKO mice is correlated with a reduced number of apoptotic cells in the wound bed. (A) Immunostaining for cleaved caspase 3 (cc3; red) and endothelial cell marker CD31 (green) on cryosections from 10-d wounds of control, α3eKO, α9eKO, and α3/α9eKO mice. Top, cleaved caspase 3 alone; bottom, overlay of cleaved caspase 3 with CD31. (B) Cleaved caspase 3 staining was quantified as the percentage of positive staining per field of 10-d wound beds from control, α3eKO, α9eKO, or α3/α9eKO mice. Means ± SEM are shown. n ≥ 5 mice per genotype. A one-way analysis of variance with Newman-Keuls’s multiple comparisons test was used. *, P

    Journal: The Journal of Cell Biology

    Article Title: Suppression of integrin α3β1 by α9β1 in the epidermis controls the paracrine resolution of wound angiogenesis

    doi: 10.1083/jcb.201510042

    Figure Lengend Snippet: Persistent vasculature in wounds of α9eKO mice is correlated with a reduced number of apoptotic cells in the wound bed. (A) Immunostaining for cleaved caspase 3 (cc3; red) and endothelial cell marker CD31 (green) on cryosections from 10-d wounds of control, α3eKO, α9eKO, and α3/α9eKO mice. Top, cleaved caspase 3 alone; bottom, overlay of cleaved caspase 3 with CD31. (B) Cleaved caspase 3 staining was quantified as the percentage of positive staining per field of 10-d wound beds from control, α3eKO, α9eKO, or α3/α9eKO mice. Means ± SEM are shown. n ≥ 5 mice per genotype. A one-way analysis of variance with Newman-Keuls’s multiple comparisons test was used. *, P

    Article Snippet: Immunohistology of tissue sections 10-µm frozen tissue sections were rehydrated in 0.2% Tween 20/PBS for 10 min, fixed in 3.7% formaldehyde, permeabilized in 0.5% Triton X-100, blocked in 10% heat-inactivated goat serum/5% milk/PBS for 1 h, and then stained with anti–cleaved caspase 3, anti-CD31 (BD), anti–LN-332 (Abcam), anti-FN (Sigma-Aldrich), anti–MMP-9 (Sigma-Aldrich), or anti–keratin 14 (Covance).

    Techniques: Mouse Assay, Immunostaining, Marker, Staining

    CPS-induced apoptosis in ALI cultures of sheep bronchial epithelial cells. Cells were pretreated with/without NAC (10 mM) for 2 h, followed by exposure to indicated conditions for 48 h. (a) 4-week-old ALI cultures of sheep bronchial epithelial cells were apically infected with CPS at 100 ng/ml and MO at MOI of 30 for 48 h before samples were harvested for analysis. Percentages of apoptotic cells were determined by flow cytometry using Annexin V/PI double-staining assay. (b) Immunoblots of apoptosis-associated proteins. The blots were probed for β -actin as a loading control. (c and d) Representative blots for cleaved-caspase-3 and cleaved-PARP1 were semiquantified by a densitometric analysis by calculating the fold of change of a protein of interest over β -actin. (e) Relative caspase-3 activity of ALI cells was detected after treating with indicated conditions. Values are mean ± SD for at least three independent experiments performed in triplicate. Compared to non-CPS or MO treatment, ∗ p

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Capsular Polysaccharide of Mycoplasma ovipneumoniae Induces Sheep Airway Epithelial Cell Apoptosis via ROS-Dependent JNK/P38 MAPK Pathways

    doi: 10.1155/2017/6175841

    Figure Lengend Snippet: CPS-induced apoptosis in ALI cultures of sheep bronchial epithelial cells. Cells were pretreated with/without NAC (10 mM) for 2 h, followed by exposure to indicated conditions for 48 h. (a) 4-week-old ALI cultures of sheep bronchial epithelial cells were apically infected with CPS at 100 ng/ml and MO at MOI of 30 for 48 h before samples were harvested for analysis. Percentages of apoptotic cells were determined by flow cytometry using Annexin V/PI double-staining assay. (b) Immunoblots of apoptosis-associated proteins. The blots were probed for β -actin as a loading control. (c and d) Representative blots for cleaved-caspase-3 and cleaved-PARP1 were semiquantified by a densitometric analysis by calculating the fold of change of a protein of interest over β -actin. (e) Relative caspase-3 activity of ALI cells was detected after treating with indicated conditions. Values are mean ± SD for at least three independent experiments performed in triplicate. Compared to non-CPS or MO treatment, ∗ p

    Article Snippet: Antibodies against Bcl-xl, Bax, Bcl-2, Cytochrome c, apoptosis inducing factor (AIF), cleaved-caspase-3, CAT, SOD2 , FADD, FAS, FASL, cleaved-PARP1, cleaved-caspase-8, ERK, and β -actin were products of Proteintech (Chicago, IL, USA).

    Techniques: Infection, Flow Cytometry, Cytometry, Double Staining, Western Blot, Activity Assay

    Tet and CNTF inhibited the expression of cleaved caspase-3 in the retinas 1 day after I/R insult. Notes: Effect of Tet ( A – C , J – L ), CNTF ( D – F , M – O ) and PBS control ( G – I , P – R ) on the expression of cleaved caspase-3 1 day after retinal ischemia. Cleaved caspase-3 is green and DAPI-labeled nuclei are blue in the merged images. Red arrows indicate green fluorescence. The ganglion cell layer, inner nuclear layer, and outer nuclear layer are indicated. Scale bar =20 μm ( A – R ). All images were captured at 40× magnification. Abbreviations: CNTF, ciliary neurotrophic factor; DAPI, 4, 6-diamidino-2-phenylindole; I/R, ischemia/reperfusion; PBS, phosphate-buffered saline; Tet, tetrandrine; GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer.

    Journal: Drug Design, Development and Therapy

    Article Title: Tetrandrine protects mouse retinal ganglion cells from ischemic injury

    doi: 10.2147/DDDT.S55407

    Figure Lengend Snippet: Tet and CNTF inhibited the expression of cleaved caspase-3 in the retinas 1 day after I/R insult. Notes: Effect of Tet ( A – C , J – L ), CNTF ( D – F , M – O ) and PBS control ( G – I , P – R ) on the expression of cleaved caspase-3 1 day after retinal ischemia. Cleaved caspase-3 is green and DAPI-labeled nuclei are blue in the merged images. Red arrows indicate green fluorescence. The ganglion cell layer, inner nuclear layer, and outer nuclear layer are indicated. Scale bar =20 μm ( A – R ). All images were captured at 40× magnification. Abbreviations: CNTF, ciliary neurotrophic factor; DAPI, 4, 6-diamidino-2-phenylindole; I/R, ischemia/reperfusion; PBS, phosphate-buffered saline; Tet, tetrandrine; GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer.

    Article Snippet: Antibodies to Bcl-2 and cleaved caspase-3 (Asp175) were purchased from Beyotime (Haimen, People’s Republic of China) and JC-10 from AAT Bioquest, Inc. (Sunnyvale, CA, USA).

    Techniques: Expressing, Labeling, Fluorescence

    Effect of TSA on docetaxel‐sensitive and docetaxel‐resistant PC a cells. ( A ) The morphological changes after 24‐hrs treatment with 0, 0.1, 0.2, 0.4 and 0.8 μM of TSA in PC 3 and PC 3/Doc cells. ( B ) Cell viability was determined by the MTT assay after treatment with different TSA concentrations for 24 hrs in PC 3 and PC 3/Doc cells. ( C ) Cell proliferation was examined by EdU incorporation assay at 16 hrs after TSA treatment, and ( D ) EdU‐positive cells were calculated. ( E ) Detection of apoptotic cells after annexin V/ PI staining by flow cytometric analysis after treatment with 0, 0.1, 0.2 and 0.4 μM of TSA for 24 hrs in PC 3/Doc cells and ( F ) the population of the apoptotic cell was calculated. ( G ) Cleaved caspase‐3 and PARP expression were examined for cells treated with TSA at various concentrations for 24 hrs. ( H ) Western blot analysis of the cleaved caspase‐3, PARP , rH 2 AX , Rad 51 and Ku70/80 in PC 3/Doc cells treated with TSA (0.4 μM) at different time‐points. ( I ) Morphological changes and ( J ) cell viability in the absence or presence of pan‐caspase inhibitor (z‐ VAD ‐fmk). PC 3/Doc cells were exposed to 10 μM z‐ VAD ‐fmk for 2 hrs prior to TSA (0.4 μM) or vehicle treatment for 24 hrs. * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Hyper‐acetylation contributes to the sensitivity of chemo‐resistant prostate cancer cells to histone deacetylase inhibitor Trichostatin A

    doi: 10.1111/jcmm.13475

    Figure Lengend Snippet: Effect of TSA on docetaxel‐sensitive and docetaxel‐resistant PC a cells. ( A ) The morphological changes after 24‐hrs treatment with 0, 0.1, 0.2, 0.4 and 0.8 μM of TSA in PC 3 and PC 3/Doc cells. ( B ) Cell viability was determined by the MTT assay after treatment with different TSA concentrations for 24 hrs in PC 3 and PC 3/Doc cells. ( C ) Cell proliferation was examined by EdU incorporation assay at 16 hrs after TSA treatment, and ( D ) EdU‐positive cells were calculated. ( E ) Detection of apoptotic cells after annexin V/ PI staining by flow cytometric analysis after treatment with 0, 0.1, 0.2 and 0.4 μM of TSA for 24 hrs in PC 3/Doc cells and ( F ) the population of the apoptotic cell was calculated. ( G ) Cleaved caspase‐3 and PARP expression were examined for cells treated with TSA at various concentrations for 24 hrs. ( H ) Western blot analysis of the cleaved caspase‐3, PARP , rH 2 AX , Rad 51 and Ku70/80 in PC 3/Doc cells treated with TSA (0.4 μM) at different time‐points. ( I ) Morphological changes and ( J ) cell viability in the absence or presence of pan‐caspase inhibitor (z‐ VAD ‐fmk). PC 3/Doc cells were exposed to 10 μM z‐ VAD ‐fmk for 2 hrs prior to TSA (0.4 μM) or vehicle treatment for 24 hrs. * P

    Article Snippet: The blots were incubated with primary antibodies against PERK, p‐PERK (Thr981), ATF4 (CREB‐2), ATF3, Bcl‐2, BAX, poly (ADP‐ribose) polymerase (PARP), HDAC1 glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) (Santa Cruz Biotechnology, Dallas, TX, USA), cleaved caspase‐3 (Epitomics, Burlingame, CA, USA), mTOR and phospho‐mTOR (Ser2448), DJ‐1, GRP78, eIF2a, phospho‐eIF2a, AKT, phospho‐AKT (Ser473), HDAC5, HDAC6 (Cell Signaling Technology, Danvers, MA, USA), 4EBP1 (Abcam, Cambridge, MA, USA), HDAC2, HDAC4 and COX4 (Proteintech, Wuhan, China) overnight at 4°C, respectively, followed by appropriate peroxidase‐conjugated secondary antibodies.

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

    TSA affects protein synthesis through modulating AKT / mTOR /4 EBP 1 signalling. ( A ) Western blot analyses of the expression differences of the AKT / mTOR /4 EBP 1 pathway between PC 3 and PC 3/Doc cells. ( B ) Analysis of the effect of TSA on proteins associated with protein synthesis by Western blot analysis. ( C , D ) The PI 3K inhibitor LY 294002 attenuated TSA ‐mediated cell death. PC 3/Doc cells were incubated with LY 294002 (20 μM) for 2 hrs prior to 0.4 μM TSA treatment. Morphological changes were observed under a light microscope, cell viability was measured by the MTT assay, and cleaved caspase‐3 and PARP expression were determined by Western blot analysis. ( E, F ) PC 3/Doc cells transfected with AKT 1‐ DN for 24 hrs prior to 0.4 μM TSA treatment. Cell viability was investigated by MTT (upper panel), and cell death was determined by Western blot analysis. * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Hyper‐acetylation contributes to the sensitivity of chemo‐resistant prostate cancer cells to histone deacetylase inhibitor Trichostatin A

    doi: 10.1111/jcmm.13475

    Figure Lengend Snippet: TSA affects protein synthesis through modulating AKT / mTOR /4 EBP 1 signalling. ( A ) Western blot analyses of the expression differences of the AKT / mTOR /4 EBP 1 pathway between PC 3 and PC 3/Doc cells. ( B ) Analysis of the effect of TSA on proteins associated with protein synthesis by Western blot analysis. ( C , D ) The PI 3K inhibitor LY 294002 attenuated TSA ‐mediated cell death. PC 3/Doc cells were incubated with LY 294002 (20 μM) for 2 hrs prior to 0.4 μM TSA treatment. Morphological changes were observed under a light microscope, cell viability was measured by the MTT assay, and cleaved caspase‐3 and PARP expression were determined by Western blot analysis. ( E, F ) PC 3/Doc cells transfected with AKT 1‐ DN for 24 hrs prior to 0.4 μM TSA treatment. Cell viability was investigated by MTT (upper panel), and cell death was determined by Western blot analysis. * P

    Article Snippet: The blots were incubated with primary antibodies against PERK, p‐PERK (Thr981), ATF4 (CREB‐2), ATF3, Bcl‐2, BAX, poly (ADP‐ribose) polymerase (PARP), HDAC1 glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) (Santa Cruz Biotechnology, Dallas, TX, USA), cleaved caspase‐3 (Epitomics, Burlingame, CA, USA), mTOR and phospho‐mTOR (Ser2448), DJ‐1, GRP78, eIF2a, phospho‐eIF2a, AKT, phospho‐AKT (Ser473), HDAC5, HDAC6 (Cell Signaling Technology, Danvers, MA, USA), 4EBP1 (Abcam, Cambridge, MA, USA), HDAC2, HDAC4 and COX4 (Proteintech, Wuhan, China) overnight at 4°C, respectively, followed by appropriate peroxidase‐conjugated secondary antibodies.

    Techniques: Western Blot, Expressing, Incubation, Light Microscopy, MTT Assay, Transfection

    The effect of a DIO regimen +/− EPA+DHA supplementation on IHC tumoral staining. A, representative photomicrographs of pathology and IHC staining of tumors for cleaved caspase-3, COX-2 and phospho-p65. B, bar graphs representing the Aperio image quantitation, scale bars indicate 100 μm, n=6 per group. Means ± SEM, statistically significant (P

    Journal: Cancer prevention research (Philadelphia, Pa.)

    Article Title: Omega-3-Acid Ethyl Esters Block the Pro-tumorigenic Effects of Obesity in Mouse Models of Postmenopausal Basal-Like and Claudin-Low Breast Cancer

    doi: 10.1158/1940-6207.CAPR-15-0018

    Figure Lengend Snippet: The effect of a DIO regimen +/− EPA+DHA supplementation on IHC tumoral staining. A, representative photomicrographs of pathology and IHC staining of tumors for cleaved caspase-3, COX-2 and phospho-p65. B, bar graphs representing the Aperio image quantitation, scale bars indicate 100 μm, n=6 per group. Means ± SEM, statistically significant (P

    Article Snippet: The influence of dietary interventions on markers of cellular apoptosis and inflammation was assessed in tumor tissues by IHC staining against cleaved caspase-3, COX-2 and phospho-p65 ( ).

    Techniques: Immunohistochemistry, Staining, Quantitation Assay