caspase 3 Search Results


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  • 99
    RayBiotech caspase 3 colorimetric assay kit
    The <t>caspase-3</t> activity in HepG2 cells treated with the leaves and root extracts (IC 50 concentration) for 24 h and 48 h (n = 3) as determined by colorimetric assay. Untreated cells were used as control for comparison. Statistical comparison was made using one-way ANOVA followed by Bonferroni (* denotes p
    Caspase 3 Colorimetric Assay Kit, supplied by RayBiotech, used in various techniques. Bioz Stars score: 99/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore caspase 3 protease
    Lower levels of activated caspase 3 activity correlate with better discrimination learning. a ) Parallel relationship between cognitive scores (errors in reversal learning) and abundance of cells expressing activated caspase 3 in each animal  b ) Caspase 3 expression is directly correlated with errors in reversal learning. Pearson analysis r = 0.76, p
    Caspase 3 Protease, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    ABclonal caspase 3
    Schematic of AS-amplified JQ1 toxicity in gastric and colon cancer cells via the mitochondrial pathway. Notes: In gastric and colon cancer cells, AS and JQ1 synergistically modulated NFATc protein family and c-Myc, which were responsible for cell viability and tumor invasiveness and metastasis. AS and JQ1 exerted synergistic cytotoxicity via upregulating Bax/Bcl-2 ratio, leading to cytochrome c release from the dysfunctional mitochondrial, the <t>caspase-3</t> activation, and the subsequent cell apoptosis. The decreased tumor invasiveness and metastasis induced by AS and JQ1 were further confirmed by the decreased protein level of MMP-9, E-cadherin, and VEGF. Pointer arrow indicates stimulation; flat arrow indicates inhibition. Abbreviations: AS, arsenic sulfide; J, JQ1; MMP, mitochondrial membrane potential; NFAT, nuclear factor of activated T-cell; VEGF, vascular endothelial growth factor.
    Caspase 3, supplied by ABclonal, used in various techniques. Bioz Stars score: 99/100, based on 105 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore caspase 3
    Immunofluorescence images of calbindin-D28K ( A , B ) and cleaved <t>caspase-3</t> ( C , D ) in control, 24 hours and 7 days after blast injury. Sagittal sections with enlarged fragments from the paramedian lobules (insets) captured at 40x magnification and quantification results are presented. Paramedial lobules double stained for calbindin-D28K (red) and caspase-3 (green) showing no colocalization of caspase-3 in Purkinje cells ( E , F ). Bar plots present quantification of: 1) a cell count for: calbindin-28k ( A ), and capsase-3 ( C ) positive cells, and 2) calbindin-28k fluorescence signal ( B ). No appreciable changes in the levels of calbindin-D28K or cleaved caspase-3 were observed between control and the injured cerebella (p > 0.05).
    Caspase 3, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1580 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc anti caspase3 casp3
    Increased apoptosis in KO myotubes. ( A ) Western blot of total lysates (top) and nuclear and mitochondrial fractions of WT and KO myotubes grown in differentiation medium for 6 to 7 d. No changes in the levels of activated (cleaved) <t>CASP3</t> products are
    Anti Caspase3 Casp3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 95 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore caspase 3 activity
    ( A ) Western immunoblot showing phospho-MAPK in Sca-1 + cells preconditioned with 100 nM IGF-1 for different time durations. Parallel experiment was performed with pre-treatment of Sca-1 + cells with 50 μM of MAPK blocker PD98059 for 30 min followed by 100 nM IGF-1 for the stipulated time durations. Western blot for p42/44 MAPK with/without PD98059 was performed on the same membrane and subjected to the same antibody incubation, detection, and exposure. The blots were later edited to align for comparison. ( B and C ) Densitometry for p42 MAPK and p44 MAPK for different durations of treatment showed the kinetics of MAPK phosphorylation over time and abrogation after PD98059 blocker treatment. ( D ) Representative western immunoblot from subfractionated cell lysate samples from preconditioning of Sca-1 + cells showing enhanced mitochondrial translocation of Cx-43 which was blocked by 50 μM PD98059 for 30 min (densitometric results are an average of three separate experiments). ( E ) Parallel experiments were performed to show that <t>caspase-3</t> activity was significantly increased in the preconditioned Sca-1 + cells pre-treated with PD98059 with concomitant reduction in cell survival as determined by LDH release assay ( F ) (the number of experiments performed, n = 3).
    Caspase 3 Activity, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1541 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    WuXi AppTec caspase3
    C23 suppressed the transcriptional activity of p53 upon DNA damage and hypoxia A. pGL3-3×p53-BS-LUC and Renilla were co-transfected in HCT116 cells combined with and without C23 shRNA. 24 hours after transfection, cells were treated with Doxorubicin (100ng/ml) or upon 1% O 2 for indicated times and the cell lysates were analyzed by lucifease assay. B. HCT116 cells with and without stable overexpressing C23 were co-transfected with pGL3-3×p53-BS-LUC and Renilla. 24 hours after transfection, cells were treated with Doxorubicin (100ng/ml) or upon 1% O 2 for indicated times and then the cell lysates were subjected to lucifease assays. C. HCT116 cells with and without stable knockdown of C23 were treated with Doxorubicin (100ng/ml) for 24 hours, then p53 target gene mRNA expression levels were analyzed by qRT-PCR analysis. D-E. HCT116 cells with and without stable overexpressing C23 (a) or stable knockdown of C23(b) were treated with Doxorubicin (100ng/ml) or mock control for 24 hours. Cell lysates were then subjected to Western blot analysis with the indicated antibodies (D) and <t>caspase3/7</t> activity analysis (E), respectively. F. (a) HCT116 cells stably expressing the control shRNA, C23 shRNA, p53 shRNA,or C23 shRNA plus p53 shRNA. (b) HCT116 cells with and without p53 knockdown were introduced with C23 respectively. Protein level of C23 and p53 were evaluated by Western blot. GAPDH served as loading control. G. HCT116 cells stably expressing the control shRNA, C23 shRNA, p53 shRNA,or C23 shRNA plus p53 shRNA (a) and HCT116 cells stably knocked down p53 were additionally introduced with C23 respectively (b). Cell number was evaluated by cell counter. The data were represented as mean±S.D. from three independent experiments. H. (a) HCT116 cells stably expressing the control shRNA, C23 shRNA, p53 shRNA,or C23 shRNA plus p53 shRNA. (b) HCT116 cells stably knocked down p53 were co-transfected with C23 cDNA respectively. Cells were then treated with 100ng/ml Doxorubicin for 24 hours. Cell viability was determined by MTT assay. The data were shown as the mean±s.d. of three independent experiments.
    Caspase3, supplied by WuXi AppTec, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc caspase 3
    Heat stress induced apoptosis and <t>caspase-3</t> activation. F98 cells were incubated at 43°C for 60 min to simulate heat stress. Culture medium was then replaced prior to further incubation of the cells for 0, 3, 6 or 12 h (R0 to R12, respectively). (A) Analysis of apoptosis was performed with flow cytometry using Annexin V-FITC/PI staining. (B) The fluorogenic substrate Ac-DEVD-AMC was applied to measure enzymatic activity of caspase-3, which was expressed relative to the control cells incubated at 37°C (set as 100%). *P
    Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 21220 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc cleaved casp3 caspase3
    Heat stress induced apoptosis and <t>caspase-3</t> activation. F98 cells were incubated at 43°C for 60 min to simulate heat stress. Culture medium was then replaced prior to further incubation of the cells for 0, 3, 6 or 12 h (R0 to R12, respectively). (A) Analysis of apoptosis was performed with flow cytometry using Annexin V-FITC/PI staining. (B) The fluorogenic substrate Ac-DEVD-AMC was applied to measure enzymatic activity of caspase-3, which was expressed relative to the control cells incubated at 37°C (set as 100%). *P
    Cleaved Casp3 Caspase3, 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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    88
    Becton Dickinson caspase 3 casp3 antibody
    Placement of the VOC, umbilical artery, and OUA connection is dose-dependent on T (A) Dot-box plot of <t>CASP3+</t> cells in allantois and overlying yolk sac (EB-6s stages); means (dashed line), SEM (error bars); n.s., not significant (Student t -Test; T + /T + vs T C /T + , P =0.8912; T + /T + vs T C /T C , P =0.3714; T C /T + vs T C /T C , P =0.2134). ( B-D ) Immunostaining for FLK-1 amongst T C genotypes. Black boxed insets, lower left: enlarged prospective VOC of black-boxed region in main panels. Black arrows (main panels) and arrowheads (insets): correctly patterned FLK-1 angioblasts within distal allantois and prospective VOC. Dashed lines, posteriormost extension of primitive streak (B-D); white asterisk, ACD (B, C). Magenta inset (D): a misplaced PECAM-1-positive (magenta arrow) vessel within the yolk sac near its junction with the allantois, the region of which is indicated by the magenta box in the main panel (see text). ( E, F ) 3D models (frontal ventral views) reconstructed from PECAM-1-immunostained nascent arterial vessels at the posterior embryonic-extraembryonic interface; dashed vertical line (E, F) marks the axial midline. Color key: blue, dorsal aortae (da); red, VOC (asterisk); yellow, umbilical artery (ua). ( G ) T C /T C mutant specimen immunostained for PECAM-1, sagittal section, lateral to the midline, equivalent to the solid horizontal line in F. White arrow indicates a vessel forming at the allantoic-yolk sac junction off the midline. ( H-K ) Examples of tissue sections used for the 3D reconstructions in (L-N). (H) PECAM-1-immunostained transverse section showing the paired da on either side of the hindgut (hg), and the omphalomesenteric artery (oa) at the ventral midline. (I-K) Colorized transverse sections from the level of the oa (H, I) and proceeding posteriorly (distally), to capture the VOC (J) and ua (K). All reconstructed sections were PECAM-1 immunostained. ( L-N ) Top, ventral (frontal) and side views (posterior, right) of the OUA connection in all three T genotypes, colored as described above with addition of purple for the oa. Red arrows (N) indicate missing OUA connection; vertical dashed line indicates the axial midline and the site where the VOC (red, located off center) should be located. Scale bar (G): 10 μm (D magenta inset); 20 μm (B-D, H-K), 25 μm (G). al, allantois; am, amnion; em, embryo; ys, yolk sac.
    Caspase 3 Casp3 Antibody, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Boster Bio anti caspase 3 casp3 picoband antibody
    Placement of the VOC, umbilical artery, and OUA connection is dose-dependent on T (A) Dot-box plot of <t>CASP3+</t> cells in allantois and overlying yolk sac (EB-6s stages); means (dashed line), SEM (error bars); n.s., not significant (Student t -Test; T + /T + vs T C /T + , P =0.8912; T + /T + vs T C /T C , P =0.3714; T C /T + vs T C /T C , P =0.2134). ( B-D ) Immunostaining for FLK-1 amongst T C genotypes. Black boxed insets, lower left: enlarged prospective VOC of black-boxed region in main panels. Black arrows (main panels) and arrowheads (insets): correctly patterned FLK-1 angioblasts within distal allantois and prospective VOC. Dashed lines, posteriormost extension of primitive streak (B-D); white asterisk, ACD (B, C). Magenta inset (D): a misplaced PECAM-1-positive (magenta arrow) vessel within the yolk sac near its junction with the allantois, the region of which is indicated by the magenta box in the main panel (see text). ( E, F ) 3D models (frontal ventral views) reconstructed from PECAM-1-immunostained nascent arterial vessels at the posterior embryonic-extraembryonic interface; dashed vertical line (E, F) marks the axial midline. Color key: blue, dorsal aortae (da); red, VOC (asterisk); yellow, umbilical artery (ua). ( G ) T C /T C mutant specimen immunostained for PECAM-1, sagittal section, lateral to the midline, equivalent to the solid horizontal line in F. White arrow indicates a vessel forming at the allantoic-yolk sac junction off the midline. ( H-K ) Examples of tissue sections used for the 3D reconstructions in (L-N). (H) PECAM-1-immunostained transverse section showing the paired da on either side of the hindgut (hg), and the omphalomesenteric artery (oa) at the ventral midline. (I-K) Colorized transverse sections from the level of the oa (H, I) and proceeding posteriorly (distally), to capture the VOC (J) and ua (K). All reconstructed sections were PECAM-1 immunostained. ( L-N ) Top, ventral (frontal) and side views (posterior, right) of the OUA connection in all three T genotypes, colored as described above with addition of purple for the oa. Red arrows (N) indicate missing OUA connection; vertical dashed line indicates the axial midline and the site where the VOC (red, located off center) should be located. Scale bar (G): 10 μm (D magenta inset); 20 μm (B-D, H-K), 25 μm (G). al, allantois; am, amnion; em, embryo; ys, yolk sac.
    Anti Caspase 3 Casp3 Picoband Antibody, supplied by Boster Bio, used in various techniques. Bioz Stars score: 99/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam caspase 3
    OLE affects Bcl-2 family, cleaved <t>caspase-3</t> protein expression, and Caspase-3 activity on day 5 after IRI or sham. Representative Western blots and densitometric quantification of Bax/Bcl-2 ( A ). Representative Western blots and densitometric quantification and cleaved caspase-3/β-actin on day 5 after IRI ( B ). Caspase-3 activity in brain tissues of rats in different groups ( C ). Data are presented as the mean ±SEM (* P
    Caspase 3, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 4059 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher caspase 3
    The mRNA ( A ) and protein ( B ) expressions of <t>caspase-3</t> caspase-8 and caspase-9 in human oral squamous carcinoma HSC-3 cells. * p
    Caspase 3, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 478 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Cell Signaling Technology Inc cleaved caspase 3 casp3
    Substitution of Matrigel/rCOL by hCOL from fibroblasts leads to lumenized EC network formation with involvement of apoptosis. ( a , b ) 3D hydrogel constructs containing GFP positive HUVECs and hASCs and hCOL after 14 days of cultivation in SFM. ( c ) Virtual stacks of GFP-HUVECs co-cultured with hASCs in hCOL for 14 days and stained for α-SMA. Nuclei were counterstained with DAPI. α-SMA positive cells are in physical contact with GFP-HUVECs. ( d ) Virtual stacks of HUVECs co-cultured with hASCs and incubated with Texas red-labeled Dextran. ( e – e ”’) Images of cryo-sections from constructs containing GFP-HUVECs, hASCs and hCOL. Constructs were cultivated for 14 days in SFM and incubated with Texas red-labeled dextran. Cryo-sections were stained with DAPI. ( e ) GFP-HUVEC, ( e ’) DAPI, ( e ”) Texas red-labeled dextran, ( e ”’) Superimposed image of ( e , e ’, e ”). Intense DAPI staining of fragmented DNA is localized within the lumen filled with Texas red-labeled dextran. ( f – f ”’) Images of cryo-sections from constructs containing GFP-HUVECs, hASCs and hCOL. Constructs were cultivated for 14 days in SFM. Cryo-sections were stained for cleaved caspase 3 and DAPI. ( f ) GFP-HUVEC, ( f ’) DAPI, ( f ”) cleaved caspase 3 <t>(CASP3),</t> ( f ”’) superimposed image of ( f , f ’, f ”). Intense DAPI staining of fragmented DNA is localized within the lumen and co-localizes with the signal for cleaved caspase 3. Scale bar: ( a ) 500 µm; ( b ) 200 µm; ( c – f ”’) 30 µm.
    Cleaved Caspase 3 Casp3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Novus Biologicals caspase 3 antibodies
    1, 25(OH)2 D3 induces apoptosis of PEL cells. PEL cells were cultured in the presence or absence of 1, 25(OH)2 D3 (10 nM). ( A ) After 48 h in culture, cells were washed and stained with Annexin V and PI and analyzed by flow cytometry. The numbers in the lower right quadrant represent the percent of apoptotic cells in culture. ( B and C ) . Cleaved <t>caspase-3,</t> and cleaved PARP expressions were detected by Western blot in JSC-1 and HBL-6 cells after treatment with 1, 25(OH)2 D3. β-actin was used to normalize protein loading.
    Caspase 3 Antibodies, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    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: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Upstate Biotechnology Inc apopain yama cpp32 caspase 3 agarose
    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
    Apopain Yama Cpp32 Caspase 3 Agarose, supplied by Upstate Biotechnology Inc, used in various techniques. Bioz Stars score: 84/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore caspase 3 casp3 antibody
    Western blot analysis of Cx43, HC1, and <t>Casp3</t> expression from astrocytes following OGD treatment. Astrocytes were treated with OGD techniques. The lysates were resolved by SDS-PAGE, and immunopositive bands were semiquantified by scanning laser densitometry. Data are expressed as mean ± SD corresponding to data displayed, * P
    Caspase 3 Casp3 Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc casp3 caspase3
    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.
    Casp3 Caspase3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore recombinant human caspase 3
    When neither CTSB deletion nor pH neutralization affected necrosis, we investigated <t>caspase</t> 3 activation and apoptosis at the time point of maximal protease activation in acini following CCK stimulation (30 min). A , pH neutralization with chloroquine
    Recombinant Human Caspase 3, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 70 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    BioVision caspase 3
    Nickel exposure induced variation of <t>caspase-3</t> (A) and caspase-9 (B) activities in piscine brain exposed to 0% (0.00 mg L -1 ), 10% (4.1 mg L -1 ) and 20% (8.2 mg L -1 ) of 96 h LC 50 of Ni at 20, 40 and 60 days of exposure. Data are mean ± SE of eight observations. Results of DMR Test have been represented by small letters. Common letter between any two bars indicates their similarity while two different letters indicate significant difference at 5% level.
    Caspase 3, supplied by BioVision, used in various techniques. Bioz Stars score: 99/100, based on 273 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc a casp3
    A and B show percent staining for Ki67 and Mcm-2 with 95% confidence intervals across progressive tissue compartments. There is a strong shift in proliferation from basal to luminal cell compartments in both markers. C and D show <t>a-casp3</t> and Bcl-2 scores with 95% confidence intervals. Only the luminal compartment was scored for these 2 markers. For a-casp3 a sharp significant drop in expression was seen in HGPIN and cancer compartments. No differential trend was observed for Bcl-2. Supernormal glands showed significantly higher luminal Mcm-2 indices and higher a-casp-3 activity than normal glands suggestive of a field effect.
    A Casp3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore cleaved casp3 caspase3
    A and B show percent staining for Ki67 and Mcm-2 with 95% confidence intervals across progressive tissue compartments. There is a strong shift in proliferation from basal to luminal cell compartments in both markers. C and D show <t>a-casp3</t> and Bcl-2 scores with 95% confidence intervals. Only the luminal compartment was scored for these 2 markers. For a-casp3 a sharp significant drop in expression was seen in HGPIN and cancer compartments. No differential trend was observed for Bcl-2. Supernormal glands showed significantly higher luminal Mcm-2 indices and higher a-casp-3 activity than normal glands suggestive of a field effect.
    Cleaved Casp3 Caspase3, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti active casp3 caspase 3 antibody
    BSO decreased cell proliferation and induced cell apoptosis in ESCC. ( a ) PCNA IHC for cell proliferation in control and BSO-treated ESCC. ( b ) Quantification of cell proliferation in control and BSO-treated ESCC. ( c ) Cleaved <t>caspase3</t> indicating cell apoptosis in control and BSO-treated ESCC. ( d ) Quantification of cell apoptosis in control and BSO-treated ESCC. IHC, immunohistochemical.
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    Novus Biologicals casp3 anti body
    BSO decreased cell proliferation and induced cell apoptosis in ESCC. ( a ) PCNA IHC for cell proliferation in control and BSO-treated ESCC. ( b ) Quantification of cell proliferation in control and BSO-treated ESCC. ( c ) Cleaved <t>caspase3</t> indicating cell apoptosis in control and BSO-treated ESCC. ( d ) Quantification of cell apoptosis in control and BSO-treated ESCC. IHC, immunohistochemical.
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    WuXi AppTec active caspase 3
    FKBP11 and GRP78 expression levels are increased in TNBS-induced colitis. Western blot analyses revealed that the expression levels of (A) FKBP11 and GRP78, and (B) active <t>caspase-3</t> in TNBS induced colitis are significantly enhanced 3 days post-treatment compared with the ETOH group. Bar graphs demonstrate the quantitative analysis of FKBP11, GRP78 and active caspase-3 vs. GAPDH. (C) Immunohistochemistry analysis of FKBP11 and GRP78 expression in colonic mucosa of mice treated with TNBS for 3 days (scale bar, 50 µm). Data are presented as mean ± standard error (n=3). *P
    Active Caspase 3, supplied by WuXi AppTec, used in various techniques. Bioz Stars score: 99/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti casp3 c8487
    FKBP11 and GRP78 expression levels are increased in TNBS-induced colitis. Western blot analyses revealed that the expression levels of (A) FKBP11 and GRP78, and (B) active <t>caspase-3</t> in TNBS induced colitis are significantly enhanced 3 days post-treatment compared with the ETOH group. Bar graphs demonstrate the quantitative analysis of FKBP11, GRP78 and active caspase-3 vs. GAPDH. (C) Immunohistochemistry analysis of FKBP11 and GRP78 expression in colonic mucosa of mice treated with TNBS for 3 days (scale bar, 50 µm). Data are presented as mean ± standard error (n=3). *P
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    Millipore casp3 kit
    FKBP11 and GRP78 expression levels are increased in TNBS-induced colitis. Western blot analyses revealed that the expression levels of (A) FKBP11 and GRP78, and (B) active <t>caspase-3</t> in TNBS induced colitis are significantly enhanced 3 days post-treatment compared with the ETOH group. Bar graphs demonstrate the quantitative analysis of FKBP11, GRP78 and active caspase-3 vs. GAPDH. (C) Immunohistochemistry analysis of FKBP11 and GRP78 expression in colonic mucosa of mice treated with TNBS for 3 days (scale bar, 50 µm). Data are presented as mean ± standard error (n=3). *P
    Casp3 Kit, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech casp3
    HOXC 13 induces apoptosis of esophageal squamous cell carcinoma ( ESCC ) cells through regulating CASP 3. A, Gene ontology pathway analysis showed that genes co‐expressed with HOXC 13 were enriched in the “transcription,” “apoptotic process” and “proliferation” pathway. B,C, Quantitative RT ‐ PCR and western blot indicated that expression of CASP 3 was significantly upregulated by knockdown of HOXC 13. D,E, Z‐ DEVD ‐ FMK , a specific <t>caspase‐3</t> inhibitor, partially reversed the inhibitory effect of sh‐ HOXC 13 on the proliferation of ECA 109 cells. F, Z‐ DEVD ‐ FMK partially reversed sh RNA ‐ HOXC 13‐induced apoptosis. G, Western blot showed that Z‐ DEVD ‐ FMK decreased the expression of CASP 3 and upregulated PARP , the enzyme digestion substrate of caspase‐3
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    Abcam cleaved casp3
    Autophagy inhibition leads to dysfunctional mitochondria and apoptosis.  a  Representative western blot of mitophagy markers from D2.0 R cells plated on BME or BME plus COL1 with or without HCQ on days 1 and 5.  b  Representative images of live D2.0 R cells transfected with the GFP-LC3 reporter and stained with MitoTracker® Red CMXRos on BME or BME plus COL1 matrices for 5 days. Scale bar is 20 µm  c  Mitochondrial (MitoTracker® Green), mitochondrial reactive oxygen species (ROS; MitoSox™) and mitochondrial membrane potential (TMRM) quantification in D2.0 R cells on BME or BME plus COL1 matrices with or without HCQ (mean ± s.e.m,  n  = 60,000 cells from 3 independent experiments. Comparisons by Mann–Whitney  U -test, two-sided. ** P  ≤ 0.01; **** P  ≤ 0.0001). NT, non-treated; HCQ-D5, hydroxychloroquine treatment beginning on day 5; MFI, mean fluorescence intensity.  d  MitoTempo is protective of ROS-induced cell death in D2.0 R cells on BME treated with HCQ. Cells were pre-treated for 5 days with 20 μM MitoTempo and subsequently exposed to 50 μM HCQ for 24 hrs. Cell viability was assessed by Cytotox Glo assay (left graph. Mean ± s.e.m,  n  = 3 wells. Comparisons by unpaired two-sided  T -test) and Caspase 3 and 7 activity (right graph. Mean ± s.e.m,  n  = 3 wells. Comparisons by unpaired two-sided  T -test). Data are representative of three independent experiments
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    Image Search Results


    The caspase-3 activity in HepG2 cells treated with the leaves and root extracts (IC 50 concentration) for 24 h and 48 h (n = 3) as determined by colorimetric assay. Untreated cells were used as control for comparison. Statistical comparison was made using one-way ANOVA followed by Bonferroni (* denotes p

    Journal: Pharmacognosy Magazine

    Article Title: In vitro Anti-Proliferative Effect of Tephrosia purpurea on Human Hepatocellular Carcinoma Cells

    doi: 10.4103/0973-1296.203981

    Figure Lengend Snippet: The caspase-3 activity in HepG2 cells treated with the leaves and root extracts (IC 50 concentration) for 24 h and 48 h (n = 3) as determined by colorimetric assay. Untreated cells were used as control for comparison. Statistical comparison was made using one-way ANOVA followed by Bonferroni (* denotes p

    Article Snippet: Measurement of caspase-3 T. purpurea extracts induced caspase-3 activity in HepG2 cells was measured by caspase-3 colorimetric assay kits, according to the manufacturer protocol (Ray Biotech Caspase-3 Colorimetric Assay Kit).

    Techniques: Activity Assay, Concentration Assay, Colorimetric Assay

    Lower levels of activated caspase 3 activity correlate with better discrimination learning. a ) Parallel relationship between cognitive scores (errors in reversal learning) and abundance of cells expressing activated caspase 3 in each animal  b ) Caspase 3 expression is directly correlated with errors in reversal learning. Pearson analysis r = 0.76, p

    Journal: PLoS ONE

    Article Title: Dietary and Behavioral Interventions Protect against Age Related Activation of Caspase Cascades in the Canine Brain

    doi: 10.1371/journal.pone.0024652

    Figure Lengend Snippet: Lower levels of activated caspase 3 activity correlate with better discrimination learning. a ) Parallel relationship between cognitive scores (errors in reversal learning) and abundance of cells expressing activated caspase 3 in each animal b ) Caspase 3 expression is directly correlated with errors in reversal learning. Pearson analysis r = 0.76, p

    Article Snippet: Specifically, wells were coated with 1 µl of active caspase-3 protease (Chemicon) and incubated with frontal cortex homogenates (1 µl, 4 ug/ul) from 4 aged beagles at 37°C for 1 hour.

    Techniques: Activity Assay, Expressing

    Double labeling with caspase 3 and neuronal or glial markers. a ) Double labeling of activated caspase 3 with the neuronal marker NeuN and b ) the glial marker GFAP revealed that the majority of cells positive for activated caspase-3 were also NeuN positive.

    Journal: PLoS ONE

    Article Title: Dietary and Behavioral Interventions Protect against Age Related Activation of Caspase Cascades in the Canine Brain

    doi: 10.1371/journal.pone.0024652

    Figure Lengend Snippet: Double labeling with caspase 3 and neuronal or glial markers. a ) Double labeling of activated caspase 3 with the neuronal marker NeuN and b ) the glial marker GFAP revealed that the majority of cells positive for activated caspase-3 were also NeuN positive.

    Article Snippet: Specifically, wells were coated with 1 µl of active caspase-3 protease (Chemicon) and incubated with frontal cortex homogenates (1 µl, 4 ug/ul) from 4 aged beagles at 37°C for 1 hour.

    Techniques: Labeling, Marker

    Activation of caspase 3 is associated with increased levels of caspase-3 cleavage products. a ) Fractin immunohistochemical staining in frontal cortices of aged dogs treated with AOX and/or ENR interventions showed significant reduction in expression of active caspase 3 in the E/A group *p

    Journal: PLoS ONE

    Article Title: Dietary and Behavioral Interventions Protect against Age Related Activation of Caspase Cascades in the Canine Brain

    doi: 10.1371/journal.pone.0024652

    Figure Lengend Snippet: Activation of caspase 3 is associated with increased levels of caspase-3 cleavage products. a ) Fractin immunohistochemical staining in frontal cortices of aged dogs treated with AOX and/or ENR interventions showed significant reduction in expression of active caspase 3 in the E/A group *p

    Article Snippet: Specifically, wells were coated with 1 µl of active caspase-3 protease (Chemicon) and incubated with frontal cortex homogenates (1 µl, 4 ug/ul) from 4 aged beagles at 37°C for 1 hour.

    Techniques: Activation Assay, Immunohistochemistry, Staining, Expressing

    Immunohistochemical staining for caspase 3 in aged canine brains. a ) Caspase 3 immunohistochemical staining in frontal cortices of aged dogs treated with AOX and/or ENR interventions showed significant reduction in expression of active caspase 3. **p

    Journal: PLoS ONE

    Article Title: Dietary and Behavioral Interventions Protect against Age Related Activation of Caspase Cascades in the Canine Brain

    doi: 10.1371/journal.pone.0024652

    Figure Lengend Snippet: Immunohistochemical staining for caspase 3 in aged canine brains. a ) Caspase 3 immunohistochemical staining in frontal cortices of aged dogs treated with AOX and/or ENR interventions showed significant reduction in expression of active caspase 3. **p

    Article Snippet: Specifically, wells were coated with 1 µl of active caspase-3 protease (Chemicon) and incubated with frontal cortex homogenates (1 µl, 4 ug/ul) from 4 aged beagles at 37°C for 1 hour.

    Techniques: Immunohistochemistry, Staining, Expressing

    Double-labeling detected by immunofluorescence revealed colocalization of fractin and active caspase 3 in several cells in the frontal cortex.

    Journal: PLoS ONE

    Article Title: Dietary and Behavioral Interventions Protect against Age Related Activation of Caspase Cascades in the Canine Brain

    doi: 10.1371/journal.pone.0024652

    Figure Lengend Snippet: Double-labeling detected by immunofluorescence revealed colocalization of fractin and active caspase 3 in several cells in the frontal cortex.

    Article Snippet: Specifically, wells were coated with 1 µl of active caspase-3 protease (Chemicon) and incubated with frontal cortex homogenates (1 µl, 4 ug/ul) from 4 aged beagles at 37°C for 1 hour.

    Techniques: Labeling, Immunofluorescence

    Induction of apoptosis following depletion of SREBP in cancer cells is restricted to lipoprotein deplete conditions.  ( A ) RPE-myrAkt-ER cells were transfected with 25 nM siRNA oligonucleotides targeting SREBP1, SREBP2 or a combination of both. After 48 hours, cells were placed in medium containing 10% FCS or 1% LPDS for a further 48 hours in the presence of 100 nM 4-OHT or solvent (ethanol). Cell viability was determined by measuring caspase 3/7 activity (Apoptosis) normalized to total protein content (SRB). Graph shows mean ± SEM of three independent experiments. ( B ) The effect of SREBP depletion on cell viability in breast cancer cells. Cells were treated and analyzed as in A. Graphs show mean ± SEM of three independent experiments. Cell lines carry different mutations in components of the PI3-kinase pathway: MCF7 (PIK3CA E545K), T47D (PIK3CA L194F), HCC1954 (PIK3CA H1047R), BT549 (PTEN null ), MDA-MB-468 (PTEN null ), MDA-MB-231 (KRAS G13D) and SKBR3 (HER2 amplification). Information on cancer gene mutations was obtained from the Wellcome Trust Sanger Institute Cancer Genome Project ( http://www.sanger.ac.uk/genetics/CGP ). ( C ) Effect of depletion of SREBP1 or SREBP2 on viability of U87 glioblastoma cells. Graph shows mean ± SEM of three independent experiments. * P

    Journal: Cancer & Metabolism

    Article Title: Sterol regulatory element binding protein-dependent regulation of lipid synthesis supports cell survival and tumor growth

    doi: 10.1186/2049-3002-1-3

    Figure Lengend Snippet: Induction of apoptosis following depletion of SREBP in cancer cells is restricted to lipoprotein deplete conditions. ( A ) RPE-myrAkt-ER cells were transfected with 25 nM siRNA oligonucleotides targeting SREBP1, SREBP2 or a combination of both. After 48 hours, cells were placed in medium containing 10% FCS or 1% LPDS for a further 48 hours in the presence of 100 nM 4-OHT or solvent (ethanol). Cell viability was determined by measuring caspase 3/7 activity (Apoptosis) normalized to total protein content (SRB). Graph shows mean ± SEM of three independent experiments. ( B ) The effect of SREBP depletion on cell viability in breast cancer cells. Cells were treated and analyzed as in A. Graphs show mean ± SEM of three independent experiments. Cell lines carry different mutations in components of the PI3-kinase pathway: MCF7 (PIK3CA E545K), T47D (PIK3CA L194F), HCC1954 (PIK3CA H1047R), BT549 (PTEN null ), MDA-MB-468 (PTEN null ), MDA-MB-231 (KRAS G13D) and SKBR3 (HER2 amplification). Information on cancer gene mutations was obtained from the Wellcome Trust Sanger Institute Cancer Genome Project ( http://www.sanger.ac.uk/genetics/CGP ). ( C ) Effect of depletion of SREBP1 or SREBP2 on viability of U87 glioblastoma cells. Graph shows mean ± SEM of three independent experiments. * P

    Article Snippet: Thapsigargin and caspase 3/7 substrate were from Calbiochem.

    Techniques: Transfection, Activity Assay, Sulforhodamine B Assay, Multiple Displacement Amplification, Amplification

    Increased caspase-3 activity is indicative of increased myocellular apoptosis and may be a mechanism contributing to cardiac dilatation in male HCM mice consuming the soy diet. Caspase-3 activity is significantly attenuated in HCM males consuming the casein diet. n = 5–13 in each group. * P

    Journal: Journal of Clinical Investigation

    Article Title: Soy diet worsens heart disease in mice

    doi: 10.1172/JCI24676

    Figure Lengend Snippet: Increased caspase-3 activity is indicative of increased myocellular apoptosis and may be a mechanism contributing to cardiac dilatation in male HCM mice consuming the soy diet. Caspase-3 activity is significantly attenuated in HCM males consuming the casein diet. n = 5–13 in each group. * P

    Article Snippet: Caspase-3 activity was determined by monitoring the rate of cleavage of a fluorogenic caspase-3 specific substrate (Acetyl-AspGluValAsp-AMC; Calbiochem).

    Techniques: Activity Assay, Mouse Assay

    miR-579-3p induces melanoma cells apoptosis and inhibit cell cycle progression. ( A ) The indicated cells were transfected with miR-579-3p or scrambled miR for 48 h and apoptosis induction was evaluated by FACS analysis. WM266 transfected as described above were subjected to Western blot analysis ( B ) and to FACS analysis to evaluate cell cycle progression and caspase 3/7 activation ( C  and  D ). ( E ) miR-579-3p was transfected in the same cells in presence or not of p21Si to evaluate cell cycle progression through FACS analysis. Error bars, ±SD,  P

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

    Article Title: miR-579-3p controls melanoma progression and resistance to target therapy

    doi: 10.1073/pnas.1607753113

    Figure Lengend Snippet: miR-579-3p induces melanoma cells apoptosis and inhibit cell cycle progression. ( A ) The indicated cells were transfected with miR-579-3p or scrambled miR for 48 h and apoptosis induction was evaluated by FACS analysis. WM266 transfected as described above were subjected to Western blot analysis ( B ) and to FACS analysis to evaluate cell cycle progression and caspase 3/7 activation ( C and D ). ( E ) miR-579-3p was transfected in the same cells in presence or not of p21Si to evaluate cell cycle progression through FACS analysis. Error bars, ±SD, P

    Article Snippet: Flow cytometric analysis to determine caspase 3/7 activity was performed according to the manufacturer’s instructions (Millipore).

    Techniques: Transfection, FACS, Western Blot, Activation Assay

    Butyrate and SAHA induced apoptosis, caspase-3 activation, and cytochrome c release in HeLa cells. HeLa cells were harvested after 2 days of treatment with indicated concentrations of butyrate or SAHA. ( A ) Cell death was measured by Hoechst dye 33342

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

    Article Title: Apoptotic and autophagic cell death induced by histone deacetylase inhibitors

    doi: 10.1073/pnas.0408345102

    Figure Lengend Snippet: Butyrate and SAHA induced apoptosis, caspase-3 activation, and cytochrome c release in HeLa cells. HeLa cells were harvested after 2 days of treatment with indicated concentrations of butyrate or SAHA. ( A ) Cell death was measured by Hoechst dye 33342

    Article Snippet: N -benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (Z-VAD-FMK) and caspase-3 fluorogenic substrate were from Calbiochem.

    Techniques: Activation Assay

    Apaf-1 is required for butyrate and SAHA-induced caspase-3 activation but not cell death. Apaf-1 wild-type (WT, filled bars) and knockout (KO, open bars) MEFs were harvested after the treatment with butyrate ( A ) or SAHA ( B ) for 2 days or 3 days, and caspase-3

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

    Article Title: Apoptotic and autophagic cell death induced by histone deacetylase inhibitors

    doi: 10.1073/pnas.0408345102

    Figure Lengend Snippet: Apaf-1 is required for butyrate and SAHA-induced caspase-3 activation but not cell death. Apaf-1 wild-type (WT, filled bars) and knockout (KO, open bars) MEFs were harvested after the treatment with butyrate ( A ) or SAHA ( B ) for 2 days or 3 days, and caspase-3

    Article Snippet: N -benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (Z-VAD-FMK) and caspase-3 fluorogenic substrate were from Calbiochem.

    Techniques: Activation Assay, Knock-Out

    Overexpression of Bcl-XL blocked butyrate- and SAHA-induced cytochrome c release and caspase-3 activation, but not cell death. HeLa cell lines stably transfected with Bcl-XL (HeLa-Bcl-XL) or vector alone (HeLa) were treated for 2 days with various concentrations

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

    Article Title: Apoptotic and autophagic cell death induced by histone deacetylase inhibitors

    doi: 10.1073/pnas.0408345102

    Figure Lengend Snippet: Overexpression of Bcl-XL blocked butyrate- and SAHA-induced cytochrome c release and caspase-3 activation, but not cell death. HeLa cell lines stably transfected with Bcl-XL (HeLa-Bcl-XL) or vector alone (HeLa) were treated for 2 days with various concentrations

    Article Snippet: N -benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (Z-VAD-FMK) and caspase-3 fluorogenic substrate were from Calbiochem.

    Techniques: Over Expression, Activation Assay, Stable Transfection, Transfection, Plasmid Preparation

    Schematic of AS-amplified JQ1 toxicity in gastric and colon cancer cells via the mitochondrial pathway. Notes: In gastric and colon cancer cells, AS and JQ1 synergistically modulated NFATc protein family and c-Myc, which were responsible for cell viability and tumor invasiveness and metastasis. AS and JQ1 exerted synergistic cytotoxicity via upregulating Bax/Bcl-2 ratio, leading to cytochrome c release from the dysfunctional mitochondrial, the caspase-3 activation, and the subsequent cell apoptosis. The decreased tumor invasiveness and metastasis induced by AS and JQ1 were further confirmed by the decreased protein level of MMP-9, E-cadherin, and VEGF. Pointer arrow indicates stimulation; flat arrow indicates inhibition. Abbreviations: AS, arsenic sulfide; J, JQ1; MMP, mitochondrial membrane potential; NFAT, nuclear factor of activated T-cell; VEGF, vascular endothelial growth factor.

    Journal: Drug Design, Development and Therapy

    Article Title: Arsenic sulfide amplifies JQ1 toxicity via mitochondrial pathway in gastric and colon cancer cells

    doi: 10.2147/DDDT.S180976

    Figure Lengend Snippet: Schematic of AS-amplified JQ1 toxicity in gastric and colon cancer cells via the mitochondrial pathway. Notes: In gastric and colon cancer cells, AS and JQ1 synergistically modulated NFATc protein family and c-Myc, which were responsible for cell viability and tumor invasiveness and metastasis. AS and JQ1 exerted synergistic cytotoxicity via upregulating Bax/Bcl-2 ratio, leading to cytochrome c release from the dysfunctional mitochondrial, the caspase-3 activation, and the subsequent cell apoptosis. The decreased tumor invasiveness and metastasis induced by AS and JQ1 were further confirmed by the decreased protein level of MMP-9, E-cadherin, and VEGF. Pointer arrow indicates stimulation; flat arrow indicates inhibition. Abbreviations: AS, arsenic sulfide; J, JQ1; MMP, mitochondrial membrane potential; NFAT, nuclear factor of activated T-cell; VEGF, vascular endothelial growth factor.

    Article Snippet: Antibodies for cytochrome C, COX IV, caspase-3, and cleaved caspase-3 were purchased from ABclonal (Wuhan, China), and anti-β-actin was obtained from Proteintech Group (Wuhan, China).

    Techniques: Amplification, Activation Assay, Inhibition

    Effects of Anwulignan on the apoptosis in the liver. Notes: ( A ) Caspase-3 and β-actin protein electrophoresis images (Western blotting). ( B ) Caspase-3/β-actin column charts (Western blotting). ( C ) Effects of Anwulignan on caspase-3 contents in the liver tissue. ## P

    Journal: Clinical Interventions in Aging

    Article Title: Protective effect of Anwulignan against D-galactose-induced hepatic injury through activating p38 MAPK–Nrf2–HO-1 pathway in mice

    doi: 10.2147/CIA.S173838

    Figure Lengend Snippet: Effects of Anwulignan on the apoptosis in the liver. Notes: ( A ) Caspase-3 and β-actin protein electrophoresis images (Western blotting). ( B ) Caspase-3/β-actin column charts (Western blotting). ( C ) Effects of Anwulignan on caspase-3 contents in the liver tissue. ## P

    Article Snippet: Anwulignan (Chengdu Pufei De Biotech Co., Ltd., Chengdu, China), sodium carboxymethyl cellulose (AR) (Shandong Weifang Lite Composite Materials Co., Ltd., Weifang, China), Twain-20 (AR) (Tianjin Yungtay Reagent Company, Tianjin, China), d -gal (Sigma-Aldrich Co., St Louis, MO, USA), polyvinylidene fluoride (PVDF) film, HCl-Tris, 30% acrylamide, N , N , N ′, N ′-tetramethylethylenediamine (TEMED), ammonium persulfate, Tris hydroxy methyl aminomethan, glycine, and diethypyrocarbonate (DEPC) water (Beijing Dinguo Reagent Company, Beijing, China), aspartate aminotransferase (AST) kit, alanine aminotransferase (ALT) kit, 8-hydroxy-2- deoxyguanosine (8-OHdG) kit, glutathione peroxidase (GSH-Px) kit, superoxide dismutase (SOD) kit, malonaldehyde (MDA) kit, and DNA marker (Nanjing Jiancheng Bioengineering Institute, Nangjing, China), skim milk powder (BD Company, San Francisco, CA, USA), rabbit anti-p38 mitogen-activated protein kinase (MAPK) antibody, rabbit anti-phospho-p38 MAPK (T180/Y182) antibody, rabbit anti-Nrf2 antibody (EPR1390Y), and rabbit anti-HO-1 antibody (EP1808Y) (Abcam, San Francisco, CA, USA), rabbit anti-caspase-3 (0206130101) (ABclonal Biotechnology Co., Ltd., Wuhan, China), caspase-3 ELISA kit (MLBIO Biotechnology Co., Ltd., Shanghai, China); electrochemiluminescence (ECL) color liquid (Biyuntian Biological Products Co., Ltd., Beijing, China), RNA extraction kit (Vazyme Biotech Co., Ltd., Nangjing, China), broad spectrum protein marker (Beijing Soledao Technology Co., Ltd., Beijing, China), agarose (Shanghai Genview Company Co., Ltd., Shanghai, China), and HepG2 cells (American Type Culture Collection [ATCC], Manassas, VA, USA) were used.

    Techniques: Protein Electrophoresis, Western Blot

    Immunofluorescence images of calbindin-D28K ( A , B ) and cleaved caspase-3 ( C , D ) in control, 24 hours and 7 days after blast injury. Sagittal sections with enlarged fragments from the paramedian lobules (insets) captured at 40x magnification and quantification results are presented. Paramedial lobules double stained for calbindin-D28K (red) and caspase-3 (green) showing no colocalization of caspase-3 in Purkinje cells ( E , F ). Bar plots present quantification of: 1) a cell count for: calbindin-28k ( A ), and capsase-3 ( C ) positive cells, and 2) calbindin-28k fluorescence signal ( B ). No appreciable changes in the levels of calbindin-D28K or cleaved caspase-3 were observed between control and the injured cerebella (p > 0.05).

    Journal: Scientific Reports

    Article Title: Electrophysiological Correlates of Blast-Wave Induced Cerebellar Injury

    doi: 10.1038/s41598-018-31728-4

    Figure Lengend Snippet: Immunofluorescence images of calbindin-D28K ( A , B ) and cleaved caspase-3 ( C , D ) in control, 24 hours and 7 days after blast injury. Sagittal sections with enlarged fragments from the paramedian lobules (insets) captured at 40x magnification and quantification results are presented. Paramedial lobules double stained for calbindin-D28K (red) and caspase-3 (green) showing no colocalization of caspase-3 in Purkinje cells ( E , F ). Bar plots present quantification of: 1) a cell count for: calbindin-28k ( A ), and capsase-3 ( C ) positive cells, and 2) calbindin-28k fluorescence signal ( B ). No appreciable changes in the levels of calbindin-D28K or cleaved caspase-3 were observed between control and the injured cerebella (p > 0.05).

    Article Snippet: We also performed a double immunostaining of calbindin D-28K (mouse monoclonal, Calbiochem, 1:400) and caspase-3 (Rabbit polyclonal, Millipore 1:100).

    Techniques: Immunofluorescence, Staining, Cell Counting, Fluorescence

    Increased apoptosis in KO myotubes. ( A ) Western blot of total lysates (top) and nuclear and mitochondrial fractions of WT and KO myotubes grown in differentiation medium for 6 to 7 d. No changes in the levels of activated (cleaved) CASP3 products are

    Journal: Autophagy

    Article Title: Defects in calcium homeostasis and mitochondria can be reversed in Pompe disease

    doi: 10.1080/15548627.2015.1009779

    Figure Lengend Snippet: Increased apoptosis in KO myotubes. ( A ) Western blot of total lysates (top) and nuclear and mitochondrial fractions of WT and KO myotubes grown in differentiation medium for 6 to 7 d. No changes in the levels of activated (cleaved) CASP3 products are

    Article Snippet: The following antibodies were used for western blots: VCL/Vinculin (Sigma, V9131), LC3B (Sigma, L8918), CACNB1 (S7–18; Abcam, ab85020), MFN2/mitofusin 2 (Abcam, ab50830), COX4I1/COXIV (Abcam, ab14744), FIS1/TTC11/ Fission 1 (Abcam, ab71498), PARK2/Parkin (Abcam, ab15954), TUBA1A/α-tubulin (Abcam, ab18251), PINK1 (Abcam, ab23707), ubiquitin (linkage-specific K63; Millipore, 05–1308), SQSTM1 (Santa Cruz Biotechnology, sc-25575), CASP3/caspase 3 (Cell Signaling Technology, 9662), AIFM1/AIF (Cell Signaling Technology, 5318), HIST1H3A/ Histone H3 (D1H2; Cell Signaling Technology, 4499), OPA1 (BD Biosciences, 612606), DNM1L/DRP1 (DNM1L; BD Biosciences, 611738), Alexa Fluor-conjugated secondary anti-mouse (Life Technologies, A-21057) or anti-rabbit (Life Technologies, A-21076).

    Techniques: Western Blot

    A : ATPAF1 (target of miR-26a, miR-28–5p, let-7i-5p, and let-7e-5p). B : BACE1 (target of miR-103–3p, miR-374–5p, miR-7a-5p and miR-19a-3p-3p). C : CASP3 (target of miR-103-rp, let-7e-5p and let-7i-5p). D : citrate synthase (CS) (target of miR-19a-3p-3p). E : GPD2 (target of miR-30a-5p). F : LRRK2 (target of miR-19a-3p-3p and miR-181a-5p). G : MAP2K4 (target of miR-24–3p and miR-374–5p). H : VDAC1 (target of miR-7a-5p) total protein content in the TA of HCR and LCR rats ( n = 9). Values are arbitrary units expressed relative to stain-free total protein loading. *Significantly different ( P

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    Article Title: Expression of microRNAs and target proteins in skeletal muscle of rats selectively bred for high and low running capacity

    doi: 10.1152/ajpendo.00043.2017

    Figure Lengend Snippet: A : ATPAF1 (target of miR-26a, miR-28–5p, let-7i-5p, and let-7e-5p). B : BACE1 (target of miR-103–3p, miR-374–5p, miR-7a-5p and miR-19a-3p-3p). C : CASP3 (target of miR-103-rp, let-7e-5p and let-7i-5p). D : citrate synthase (CS) (target of miR-19a-3p-3p). E : GPD2 (target of miR-30a-5p). F : LRRK2 (target of miR-19a-3p-3p and miR-181a-5p). G : MAP2K4 (target of miR-24–3p and miR-374–5p). H : VDAC1 (target of miR-7a-5p) total protein content in the TA of HCR and LCR rats ( n = 9). Values are arbitrary units expressed relative to stain-free total protein loading. *Significantly different ( P

    Article Snippet: Primary antibodies used were polyclonal caspase-3 (CASP3) (no. 9662), leucine-rich repeat kinase 2 (LRRK2) (no. 5559) (Cell Signaling, Beverly, MA), polyclonal ATP synthase mitochondrial F1 complex assembly factor 1 (ATPAF1) (no. ab101518), beta-site APP cleaving enzyme 1 (BACE1; ab2077), citrate synthase (CS; ab96600), monoclonal glycerol-3-phosphate dehydrogenase 2 (GPD2; ab188585), MAP2K4 (ab33912), and VDAC1 (ab14734; Abcam, Cambridge, UK).

    Techniques: Staining

    BMSCs-IGF-1 resistance to hypoxia is dependent on AKT. BMSCs-NC or BMSCs-IGF-1 were treated with 0.5 μM LY294002 or an equal volume of DMSO for 24 h, and then, the cells were exposed to hypoxia for 48 h. a Cell proliferation was determined by MTS assay. b , c Cell migration was determined by Transwell assay. d Expression of p-AKT, AKT, SFRP2, β-catenin, cyclin D1, c-myc, BAX, BCL-2, and cleaved caspase-3 were determined by Western blotting. All assays were performed in triplicate (* P

    Journal: Stem Cell Research & Therapy

    Article Title: IGF-1 enhances BMSC viability, migration, and anti-apoptosis in myocardial infarction via secreted frizzled-related protein 2 pathway

    doi: 10.1186/s13287-019-1544-y

    Figure Lengend Snippet: BMSCs-IGF-1 resistance to hypoxia is dependent on AKT. BMSCs-NC or BMSCs-IGF-1 were treated with 0.5 μM LY294002 or an equal volume of DMSO for 24 h, and then, the cells were exposed to hypoxia for 48 h. a Cell proliferation was determined by MTS assay. b , c Cell migration was determined by Transwell assay. d Expression of p-AKT, AKT, SFRP2, β-catenin, cyclin D1, c-myc, BAX, BCL-2, and cleaved caspase-3 were determined by Western blotting. All assays were performed in triplicate (* P

    Article Snippet: The membranes were then blocked with 5% BSA in TBST buffer for 1 h at room temperature, and incubated with a primary antibody against caspase-3 (CST, 9662s), BAX (CST, 2774), BCL-2 (Abcam,196495), OCT4 (Boster,bs-0830R), p-AKT (CST, 13038S), AKT (CAT, 2920), ERK1/2 (CST, 9102), p-ERK1/2 (CST, 4370S), p-β-catenin (ser33/37/T41, CST, 9561s), β-catenin (Santa, sc7199), SFRP2 (Abcam, ab86329), and NFAT4 (Abcam, ab93628) at 4 °C overnight.

    Techniques: MTS Assay, Migration, Transwell Assay, Expressing, Western Blot

    Effect of transplantation of BMSCs-IGF-1 following MI. a One week after coronary ligation, BMSCs-NC or BMSCs-IGF-1 (1 × 10 6 ) were injected via the tail vein. Three weeks later, the fibrosis region (blue) was determined by Masson’s trichrome stain and the protein levels of BAX, BCL-2, and caspase-3 were determined by immunohistochemistry. b Expression of BAX, BCL-2, and cleaved caspase-3 were determined by Western blotting (* P

    Journal: Stem Cell Research & Therapy

    Article Title: IGF-1 enhances BMSC viability, migration, and anti-apoptosis in myocardial infarction via secreted frizzled-related protein 2 pathway

    doi: 10.1186/s13287-019-1544-y

    Figure Lengend Snippet: Effect of transplantation of BMSCs-IGF-1 following MI. a One week after coronary ligation, BMSCs-NC or BMSCs-IGF-1 (1 × 10 6 ) were injected via the tail vein. Three weeks later, the fibrosis region (blue) was determined by Masson’s trichrome stain and the protein levels of BAX, BCL-2, and caspase-3 were determined by immunohistochemistry. b Expression of BAX, BCL-2, and cleaved caspase-3 were determined by Western blotting (* P

    Article Snippet: The membranes were then blocked with 5% BSA in TBST buffer for 1 h at room temperature, and incubated with a primary antibody against caspase-3 (CST, 9662s), BAX (CST, 2774), BCL-2 (Abcam,196495), OCT4 (Boster,bs-0830R), p-AKT (CST, 13038S), AKT (CAT, 2920), ERK1/2 (CST, 9102), p-ERK1/2 (CST, 4370S), p-β-catenin (ser33/37/T41, CST, 9561s), β-catenin (Santa, sc7199), SFRP2 (Abcam, ab86329), and NFAT4 (Abcam, ab93628) at 4 °C overnight.

    Techniques: Transplantation Assay, Ligation, Injection, Staining, Immunohistochemistry, Expressing, Western Blot

    Effect of IGF-1 overexpression on BMSCs. a Identification of BMSCs by flow cytometry analysis. b Supernatants from cultured BMSCs-NC and BMSCs-IGF-1 were collected and subjected to ELISA to determine IGF-1 levels. c Cells were exposed to hypoxia for 48 h, and cell proliferation was determined by MTS assay. d Apoptosis was determined by TUNEL assay. e Cell migration was determined by Transwell assay. f Expression of OCT4, NANOG, cleaved caspase-3, BAX, and BCL-2 was determined by Western blotting. All assays were performed in triplicate (* P

    Journal: Stem Cell Research & Therapy

    Article Title: IGF-1 enhances BMSC viability, migration, and anti-apoptosis in myocardial infarction via secreted frizzled-related protein 2 pathway

    doi: 10.1186/s13287-019-1544-y

    Figure Lengend Snippet: Effect of IGF-1 overexpression on BMSCs. a Identification of BMSCs by flow cytometry analysis. b Supernatants from cultured BMSCs-NC and BMSCs-IGF-1 were collected and subjected to ELISA to determine IGF-1 levels. c Cells were exposed to hypoxia for 48 h, and cell proliferation was determined by MTS assay. d Apoptosis was determined by TUNEL assay. e Cell migration was determined by Transwell assay. f Expression of OCT4, NANOG, cleaved caspase-3, BAX, and BCL-2 was determined by Western blotting. All assays were performed in triplicate (* P

    Article Snippet: The membranes were then blocked with 5% BSA in TBST buffer for 1 h at room temperature, and incubated with a primary antibody against caspase-3 (CST, 9662s), BAX (CST, 2774), BCL-2 (Abcam,196495), OCT4 (Boster,bs-0830R), p-AKT (CST, 13038S), AKT (CAT, 2920), ERK1/2 (CST, 9102), p-ERK1/2 (CST, 4370S), p-β-catenin (ser33/37/T41, CST, 9561s), β-catenin (Santa, sc7199), SFRP2 (Abcam, ab86329), and NFAT4 (Abcam, ab93628) at 4 °C overnight.

    Techniques: Over Expression, Flow Cytometry, Cytometry, Cell Culture, Enzyme-linked Immunosorbent Assay, MTS Assay, TUNEL Assay, Migration, Transwell Assay, Expressing, Western Blot

    BMSCs-IGF-1 protected H9C2 rat cardiomyoblast cells against hypoxia. H9C2 were co-cultured with BMSCs-NC, BMSCs-IGF-1, or control medium, and then exposed to hypoxia for 48 h. a Cell proliferation of H9C2 was determined by MTS assay. b , c Apoptosis of H9C2 was determined by Annexin-V-FITC staining. d Expression of BCL-2, BAX, and cleaved caspase-3 in H9C2 was determined by Western blotting. All assays were performed in triplicate (* P

    Journal: Stem Cell Research & Therapy

    Article Title: IGF-1 enhances BMSC viability, migration, and anti-apoptosis in myocardial infarction via secreted frizzled-related protein 2 pathway

    doi: 10.1186/s13287-019-1544-y

    Figure Lengend Snippet: BMSCs-IGF-1 protected H9C2 rat cardiomyoblast cells against hypoxia. H9C2 were co-cultured with BMSCs-NC, BMSCs-IGF-1, or control medium, and then exposed to hypoxia for 48 h. a Cell proliferation of H9C2 was determined by MTS assay. b , c Apoptosis of H9C2 was determined by Annexin-V-FITC staining. d Expression of BCL-2, BAX, and cleaved caspase-3 in H9C2 was determined by Western blotting. All assays were performed in triplicate (* P

    Article Snippet: The membranes were then blocked with 5% BSA in TBST buffer for 1 h at room temperature, and incubated with a primary antibody against caspase-3 (CST, 9662s), BAX (CST, 2774), BCL-2 (Abcam,196495), OCT4 (Boster,bs-0830R), p-AKT (CST, 13038S), AKT (CAT, 2920), ERK1/2 (CST, 9102), p-ERK1/2 (CST, 4370S), p-β-catenin (ser33/37/T41, CST, 9561s), β-catenin (Santa, sc7199), SFRP2 (Abcam, ab86329), and NFAT4 (Abcam, ab93628) at 4 °C overnight.

    Techniques: Cell Culture, MTS Assay, Staining, Expressing, Western Blot

    PCDH18 suppression promoted cell migration and proliferation in colonic NCM460 cells through the Wnt/β-catenin signaling pathway. ( A ) PCDH18 mRNA and protein expression was efficiently inhibited in NCM460 cells transfected with siPCDH18. GAPDH was used as an internal control. ( B ) Representative colony formation assays in NCM460 cells transfected with siRNA-PCDH18 (siPCDH18) and siRNA-negative control (siNC). ( C ) Representative transwell assays in NCM460 cells transfected with siPCDH18 and siNC. ( D ) Cell cycle distribution of NCM460 cells transfected with siPCDH18 and siNC was detected by flow cytometry analysis. ( E ) Molecular interaction networks involving PCDH18 were visualized by Cytoscape3.3.0 software from different species. Blue node represents gene from Homo species. Red node represents gene from Mus species. Green node represents gene from Bos taurus species. ( F ) Protein expression levels of nonphospho (active)-β-catenin, phospho-β-catenin, total-β-catenin, phospho-GSK-3β, GSK-3β, p21, cyclin A1, cyclin D1, cyclin E1 and caspase3 in NCM460 cells transfected with siPCDH18 and siNC. ( G ) Representative images of β-catenin staining in NCM460 cells transfected with siPCDH18 and siNC.

    Journal: Scientific Reports

    Article Title: PCDH18 is frequently inactivated by promoter methylation in colorectal cancer

    doi: 10.1038/s41598-017-03133-w

    Figure Lengend Snippet: PCDH18 suppression promoted cell migration and proliferation in colonic NCM460 cells through the Wnt/β-catenin signaling pathway. ( A ) PCDH18 mRNA and protein expression was efficiently inhibited in NCM460 cells transfected with siPCDH18. GAPDH was used as an internal control. ( B ) Representative colony formation assays in NCM460 cells transfected with siRNA-PCDH18 (siPCDH18) and siRNA-negative control (siNC). ( C ) Representative transwell assays in NCM460 cells transfected with siPCDH18 and siNC. ( D ) Cell cycle distribution of NCM460 cells transfected with siPCDH18 and siNC was detected by flow cytometry analysis. ( E ) Molecular interaction networks involving PCDH18 were visualized by Cytoscape3.3.0 software from different species. Blue node represents gene from Homo species. Red node represents gene from Mus species. Green node represents gene from Bos taurus species. ( F ) Protein expression levels of nonphospho (active)-β-catenin, phospho-β-catenin, total-β-catenin, phospho-GSK-3β, GSK-3β, p21, cyclin A1, cyclin D1, cyclin E1 and caspase3 in NCM460 cells transfected with siPCDH18 and siNC. ( G ) Representative images of β-catenin staining in NCM460 cells transfected with siPCDH18 and siNC.

    Article Snippet: The membranes were incubated with the anti-PCDH18 (Santa Cruz cat#sc-104574), anti-nonphospho (active)-β-catenin, anti-phospho-β-catenin, anti-β-catenin, anti-phospho-GSK-3β, anti-GSK-3β, anti-caspase3 (Cell Signaling Technology cat#8814, 4176, 8480, 9331, 9315, 9662), anti-p21, anti-cyclin A1, anti-cyclin D1, anti-cyclin E1 (Bioworld cat#AP0713, BS1084, BS2436, BS1085) and anti-GAPDH (Kangchen cat#KC5G4) overnight at 4 °C.

    Techniques: Migration, Expressing, Transfection, Negative Control, Flow Cytometry, Cytometry, Software, Staining

    The effect of siRNA- HMGN5 on the expression of apoptosis associated proteins in LNCaP cells. LNCaP cells were infected with siRNA- HMGN5 , and representative blots are shown of ( a ) Bcl-2, Bcl-xl, Bax, Bid and ( b ) Caspase3, PARP, ENDO G and AIF. β-actin

    Journal: Asian Journal of Andrology

    Article Title: Small interfering RNA targeting HMGN5 induces apoptosis via modulation of a mitochondrial pathway and Bcl-2 family proteins in prostate cancer cells

    doi: 10.1038/aja.2012.18

    Figure Lengend Snippet: The effect of siRNA- HMGN5 on the expression of apoptosis associated proteins in LNCaP cells. LNCaP cells were infected with siRNA- HMGN5 , and representative blots are shown of ( a ) Bcl-2, Bcl-xl, Bax, Bid and ( b ) Caspase3, PARP, ENDO G and AIF. β-actin

    Article Snippet: Anti-Bcl-2, Bcl-xl, Bax, Bid, PARP, AIF, ENDO G and caspase3 antibodies (Cell Signal Tech, Beverly, MA, USA) and an anti-β-actin polyclonal antibody (sc-1616-R; Santa Cruz Biotechnology, Santa Cruz, CA, USA) were used as primary antibodies.

    Techniques: Expressing, Infection

    siRNA- HMGN5 activated caspase activity in LNCaP cells. The LNCaP cell protein extracts were analyzed for caspase3 levels, which were represented as absorbance values. Each value is mean ± SEM of six independent observations (* P

    Journal: Asian Journal of Andrology

    Article Title: Small interfering RNA targeting HMGN5 induces apoptosis via modulation of a mitochondrial pathway and Bcl-2 family proteins in prostate cancer cells

    doi: 10.1038/aja.2012.18

    Figure Lengend Snippet: siRNA- HMGN5 activated caspase activity in LNCaP cells. The LNCaP cell protein extracts were analyzed for caspase3 levels, which were represented as absorbance values. Each value is mean ± SEM of six independent observations (* P

    Article Snippet: Anti-Bcl-2, Bcl-xl, Bax, Bid, PARP, AIF, ENDO G and caspase3 antibodies (Cell Signal Tech, Beverly, MA, USA) and an anti-β-actin polyclonal antibody (sc-1616-R; Santa Cruz Biotechnology, Santa Cruz, CA, USA) were used as primary antibodies.

    Techniques: Activity Assay

    Neonatal ischemia triggers activation of caspase-3 (casp3) in both hemispheres. Cleaved casp3 immunolabeling (red) and TUNEL assay (DNA fragmentation, green) in sections of brain at the level of the dorsal hippocampus (bregma −3.3 mm) from contralateral (CL-M1, A–C ) and ipsilateral (IL-M1, D–F ) hemispheres at 48 hours postischemia (model M1). Note that active casp3 was cytosolic in the CL cortex (enlarged panel in C ), whereas it was nuclear and cytosolic and associated with TUNEL staining in the IL cortex (enlarged panel in F ). Scale bar represents 50 and 20 μm in enlarged panels. G: Spatial distribution of cleaved casp3- (red) and TUNEL-positive (green) cells and lesion area (gray) at 48 hours after ischemic injury in P7 rats in both IL and CL hemispheres. H: Representative Western blots probed with anti-active casp3 (17 kDa; Cell Signaling Technology) for protein samples isolated from the cytosolic (S2) and nuclear (P1) fractions of IL and CL cortex from ischemic rats sacrificed 48 hours after reperfusion and from sham (Sh) brain. The cytosolic marker β-actin was used as protein loading control. Note that p17 was present in the cytosolic fractions in both IL and CL tissues. In contrast, p17 was only present in the nuclear fraction for IL tissues; Fr, fraction. I: Quantification of casp3-positive cells at 4, 12, 48, and 72 hours postischemia in both IL and CL cortex.

    Journal: The American Journal of Pathology

    Article Title: Unilateral Blood Flow Decrease Induces Bilateral and Symmetric Responses in the Immature Brain

    doi: 10.2353/ajpath.2009.090257

    Figure Lengend Snippet: Neonatal ischemia triggers activation of caspase-3 (casp3) in both hemispheres. Cleaved casp3 immunolabeling (red) and TUNEL assay (DNA fragmentation, green) in sections of brain at the level of the dorsal hippocampus (bregma −3.3 mm) from contralateral (CL-M1, A–C ) and ipsilateral (IL-M1, D–F ) hemispheres at 48 hours postischemia (model M1). Note that active casp3 was cytosolic in the CL cortex (enlarged panel in C ), whereas it was nuclear and cytosolic and associated with TUNEL staining in the IL cortex (enlarged panel in F ). Scale bar represents 50 and 20 μm in enlarged panels. G: Spatial distribution of cleaved casp3- (red) and TUNEL-positive (green) cells and lesion area (gray) at 48 hours after ischemic injury in P7 rats in both IL and CL hemispheres. H: Representative Western blots probed with anti-active casp3 (17 kDa; Cell Signaling Technology) for protein samples isolated from the cytosolic (S2) and nuclear (P1) fractions of IL and CL cortex from ischemic rats sacrificed 48 hours after reperfusion and from sham (Sh) brain. The cytosolic marker β-actin was used as protein loading control. Note that p17 was present in the cytosolic fractions in both IL and CL tissues. In contrast, p17 was only present in the nuclear fraction for IL tissues; Fr, fraction. I: Quantification of casp3-positive cells at 4, 12, 48, and 72 hours postischemia in both IL and CL cortex.

    Article Snippet: Immunohistochemistry was performed on cryostat coronal sections (at 4, 12, 24 (M1), 48 (M1 to M4) and 72 (M2) hours after injury) as previously described, using anti-cleaved caspase-3 (casp3) (Asp175; Cell Signaling Technology, Ozyme, St-Quentin-en-Yvelines, France).

    Techniques: Activation Assay, Immunolabeling, TUNEL Assay, Staining, Western Blot, Isolation, Marker

    Unilateral transient CCA occlusion mainly triggers casp3 cleavage in neurons and undifferentiated cells in the P7 rat brain. Contralateral cortex from uni-CCAo (model M2) and MCAo + tCCAo (model M1) animals contain most cleaved casp3 colocalized with neurons (NeuN) ( A , B ), nestin- ( C , D ), and vimentin ( E , F ) positive cells Arrows indicate location magnified in insets . Co-localization of cleaved casp3 was observed in GABA-immunostained neurons in cortical layer III (ly III in G , and enlarged panels in H , indicated by the asterisk in G ). Cleaved casp3 labeling is shown in red, whereas NeuN, nestin, vimentin and GABA markers are shown in green. Scale bar represents 50 and 20 μm (enlarged panels).

    Journal: The American Journal of Pathology

    Article Title: Unilateral Blood Flow Decrease Induces Bilateral and Symmetric Responses in the Immature Brain

    doi: 10.2353/ajpath.2009.090257

    Figure Lengend Snippet: Unilateral transient CCA occlusion mainly triggers casp3 cleavage in neurons and undifferentiated cells in the P7 rat brain. Contralateral cortex from uni-CCAo (model M2) and MCAo + tCCAo (model M1) animals contain most cleaved casp3 colocalized with neurons (NeuN) ( A , B ), nestin- ( C , D ), and vimentin ( E , F ) positive cells Arrows indicate location magnified in insets . Co-localization of cleaved casp3 was observed in GABA-immunostained neurons in cortical layer III (ly III in G , and enlarged panels in H , indicated by the asterisk in G ). Cleaved casp3 labeling is shown in red, whereas NeuN, nestin, vimentin and GABA markers are shown in green. Scale bar represents 50 and 20 μm (enlarged panels).

    Article Snippet: Immunohistochemistry was performed on cryostat coronal sections (at 4, 12, 24 (M1), 48 (M1 to M4) and 72 (M2) hours after injury) as previously described, using anti-cleaved caspase-3 (casp3) (Asp175; Cell Signaling Technology, Ozyme, St-Quentin-en-Yvelines, France).

    Techniques: Labeling

    Casp3 cleavage and DNA fragmentation in several models of carotid occlusion at 48 hours after injury. Spatial distribution ( A , E , and I ) of cleaved casp3 (red), TUNEL-positive (green) cells, and lesion area (gray) after uni-CCAo (model M2), bi-tCCAo (model M3) and bi-pCCAo (model M4), respectively. Note that images in mirror were detected in all three ischemic conditions. Double fluorescent staining in the CL cortex for cleaved casp3 ( B , F , and J ) and TUNEL ( C , top , G , and K ) or Fluorojade B (FluJB) labeling ( C , bottom ). After uni-CCAo, only cytosolic cleaved casp3 (enlarged image in D ) without TUNEL or FluJB staining was observed. In contrast, several cells displayed cytosolic cleaved casp3 co-localized with TUNEL staining ( F–H , note chromatin clumps typical of apoptotic cells, enlarged panel in H ) after model M3. A large number of cells exhibited both cytosolic and nuclear cleaved casp3 and DNA fragmentation after model M4 ( J–L ). Some sections were counterstained with 4′,6′-diamidino-2-phenylindole (overlay in D and L ). Scale bar represents 50 and 20 μm (enlarged panels).

    Journal: The American Journal of Pathology

    Article Title: Unilateral Blood Flow Decrease Induces Bilateral and Symmetric Responses in the Immature Brain

    doi: 10.2353/ajpath.2009.090257

    Figure Lengend Snippet: Casp3 cleavage and DNA fragmentation in several models of carotid occlusion at 48 hours after injury. Spatial distribution ( A , E , and I ) of cleaved casp3 (red), TUNEL-positive (green) cells, and lesion area (gray) after uni-CCAo (model M2), bi-tCCAo (model M3) and bi-pCCAo (model M4), respectively. Note that images in mirror were detected in all three ischemic conditions. Double fluorescent staining in the CL cortex for cleaved casp3 ( B , F , and J ) and TUNEL ( C , top , G , and K ) or Fluorojade B (FluJB) labeling ( C , bottom ). After uni-CCAo, only cytosolic cleaved casp3 (enlarged image in D ) without TUNEL or FluJB staining was observed. In contrast, several cells displayed cytosolic cleaved casp3 co-localized with TUNEL staining ( F–H , note chromatin clumps typical of apoptotic cells, enlarged panel in H ) after model M3. A large number of cells exhibited both cytosolic and nuclear cleaved casp3 and DNA fragmentation after model M4 ( J–L ). Some sections were counterstained with 4′,6′-diamidino-2-phenylindole (overlay in D and L ). Scale bar represents 50 and 20 μm (enlarged panels).

    Article Snippet: Immunohistochemistry was performed on cryostat coronal sections (at 4, 12, 24 (M1), 48 (M1 to M4) and 72 (M2) hours after injury) as previously described, using anti-cleaved caspase-3 (casp3) (Asp175; Cell Signaling Technology, Ozyme, St-Quentin-en-Yvelines, France).

    Techniques: TUNEL Assay, Staining, Labeling

    Unilateral transient CCA occlusion can induce casp3 cleavage but not its downstream subtracts. A: Representative Western blots showing cleaved casp3 fragment (17 kDa) both in the IL and CL 48 hours after uni-tCCAo + MCAo (model M1) and after uni-CCAo (model M2) as compared with sham-operated neocortical cytosolic extracts ( n = 8 for each condition). Highly cleaved casp3 was present in the IL after model M1, but it also is increased in the CL cortex and after model M2 compared with sham (sh) cytosolic extracts ( P

    Journal: The American Journal of Pathology

    Article Title: Unilateral Blood Flow Decrease Induces Bilateral and Symmetric Responses in the Immature Brain

    doi: 10.2353/ajpath.2009.090257

    Figure Lengend Snippet: Unilateral transient CCA occlusion can induce casp3 cleavage but not its downstream subtracts. A: Representative Western blots showing cleaved casp3 fragment (17 kDa) both in the IL and CL 48 hours after uni-tCCAo + MCAo (model M1) and after uni-CCAo (model M2) as compared with sham-operated neocortical cytosolic extracts ( n = 8 for each condition). Highly cleaved casp3 was present in the IL after model M1, but it also is increased in the CL cortex and after model M2 compared with sham (sh) cytosolic extracts ( P

    Article Snippet: Immunohistochemistry was performed on cryostat coronal sections (at 4, 12, 24 (M1), 48 (M1 to M4) and 72 (M2) hours after injury) as previously described, using anti-cleaved caspase-3 (casp3) (Asp175; Cell Signaling Technology, Ozyme, St-Quentin-en-Yvelines, France).

    Techniques: Western Blot

    Molecular analysis with representative images of active caspase-3 immunostaining of CT ( a,d ), SF ( b,e ) and VTo ( c,f ) ovaries cultured ( d–f ) or not ( a–c ) for 4 hours. Illustration of labelled antral follicles ( g ) with more ( h ) or less ( i ) than 30% of active caspase-3 positive granulosa cells. ( j ) Computer-assisted quantification of active caspase-3 immunostaining. ( k ) Bax/Bcl-xl mRNA ratios and ( l ) Becn1 mRNA levels in the different groups. ( m ) Western blot quantification of cleaved caspase-3 and ( n ) LC3 protein level in the different groups. Results are expressed as means ± SEM. n = 4–16 for ( a–j ); n = 9–22 for ( k,l ); n = 5–16 for m and n per group. CT: control/fresh ovaries; SF: slow frozen/thawed ovaries, VTo: vitrified/warmed ovaries, cult: ex vivo culture during 4 hours. ns: non-significant, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, a different from CT-cult, b different from SF-cult and c different from CT.

    Journal: Scientific Reports

    Article Title: Slow Freezing Versus Vitrification of Mouse Ovaries: from Ex Vivo Analyses to Successful Pregnancies after Auto-Transplantation

    doi: 10.1038/s41598-019-56182-8

    Figure Lengend Snippet: Molecular analysis with representative images of active caspase-3 immunostaining of CT ( a,d ), SF ( b,e ) and VTo ( c,f ) ovaries cultured ( d–f ) or not ( a–c ) for 4 hours. Illustration of labelled antral follicles ( g ) with more ( h ) or less ( i ) than 30% of active caspase-3 positive granulosa cells. ( j ) Computer-assisted quantification of active caspase-3 immunostaining. ( k ) Bax/Bcl-xl mRNA ratios and ( l ) Becn1 mRNA levels in the different groups. ( m ) Western blot quantification of cleaved caspase-3 and ( n ) LC3 protein level in the different groups. Results are expressed as means ± SEM. n = 4–16 for ( a–j ); n = 9–22 for ( k,l ); n = 5–16 for m and n per group. CT: control/fresh ovaries; SF: slow frozen/thawed ovaries, VTo: vitrified/warmed ovaries, cult: ex vivo culture during 4 hours. ns: non-significant, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, a different from CT-cult, b different from SF-cult and c different from CT.

    Article Snippet: Anti-cleaved caspase-3 antibody (Cell Signaling, Danvers, USA) was diluted 1/300 in the REAL antibody diluent (Dako, Glostrup, Denmark) and incubated overnight at 4 °C, followed by incubation with the secondary antibody HRP linked (ENVISION/HRP ready to use, Dako, Glostrup, Denmark) for 30 min at RT.

    Techniques: Immunostaining, Cell Culture, Western Blot, Ex Vivo

    An in vivo evaluation of 1-MT+TL/DCs-TL vaccination efficacy in C57BL/6 black mice challenged Pan02 tumor model showed that1-MT and 1-MT+TL/DCs-TL significantly reduced tumor size 2.144 cm 3 , 1.686 cm 3 , 2.166 cm 3 respectively ( A ), tumor volume and weight also significantly reduced ( C , D ). Besides survival rate of 1-MT+TL/1-MT+DCs-TL treated groups was significantly improved ( B ), 38.64%, and 30.77% respectively in comparison with PBS group. Further Haematoxylin and Eosin (H E) staining showed that, mice treated by 1-MT and 1-MT+TL ( E .a,b) significantly represented many focal necrosis accompanied with more inflammatory infiltrating cells (black arrow for focal necrosis, and red arrow for inflammatory cells). Meanwhile 1-MT+DC-TL treated group ( E .c) showed small tumor necrosis and few numbers of inflammatory infiltrating cells. Other tested groups ( E .d,e,f and g) represented a small focal necrosis inside tumor tissue and a little number of inflammatory cells. The assessment of tumor fibrosis showed that 1-MT, 1-MT+TL, and 1-MT+DCs-TL groups extremely decreased collagen deposition folding rate in treated tumors as shown around pointed area ( F .a,b,c), while other groups showed high collagen fibers ( F .d,e,f,g). Besides that 1-MT treated groups showed significant immune reactivity of cleavage caspase3 pathway ( G .a,b,c) that indicated promoting of tumor apoptosis. While other untreated groups showed no significant apoptotic reactions ( G .d,e,f,g). These results indicated that administration of 1-Methyl-D-tryptophan and TAA vaccine destroyed tumor stromal and fibrosis, while it promoted significant apoptosis and cell death, which is very interesting observation. ***( P

    Journal: Scientific Reports

    Article Title: 1-Methyl-D-tryptophan Reduces Tumor CD133+ cells, Wnt/β-catenin and NF-κβp65 while Enhances Lymphocytes NF-κβ2, STAT3, and STAT4 Pathways in Murine Pancreatic Adenocarcinoma

    doi: 10.1038/s41598-018-28238-8

    Figure Lengend Snippet: An in vivo evaluation of 1-MT+TL/DCs-TL vaccination efficacy in C57BL/6 black mice challenged Pan02 tumor model showed that1-MT and 1-MT+TL/DCs-TL significantly reduced tumor size 2.144 cm 3 , 1.686 cm 3 , 2.166 cm 3 respectively ( A ), tumor volume and weight also significantly reduced ( C , D ). Besides survival rate of 1-MT+TL/1-MT+DCs-TL treated groups was significantly improved ( B ), 38.64%, and 30.77% respectively in comparison with PBS group. Further Haematoxylin and Eosin (H E) staining showed that, mice treated by 1-MT and 1-MT+TL ( E .a,b) significantly represented many focal necrosis accompanied with more inflammatory infiltrating cells (black arrow for focal necrosis, and red arrow for inflammatory cells). Meanwhile 1-MT+DC-TL treated group ( E .c) showed small tumor necrosis and few numbers of inflammatory infiltrating cells. Other tested groups ( E .d,e,f and g) represented a small focal necrosis inside tumor tissue and a little number of inflammatory cells. The assessment of tumor fibrosis showed that 1-MT, 1-MT+TL, and 1-MT+DCs-TL groups extremely decreased collagen deposition folding rate in treated tumors as shown around pointed area ( F .a,b,c), while other groups showed high collagen fibers ( F .d,e,f,g). Besides that 1-MT treated groups showed significant immune reactivity of cleavage caspase3 pathway ( G .a,b,c) that indicated promoting of tumor apoptosis. While other untreated groups showed no significant apoptotic reactions ( G .d,e,f,g). These results indicated that administration of 1-Methyl-D-tryptophan and TAA vaccine destroyed tumor stromal and fibrosis, while it promoted significant apoptosis and cell death, which is very interesting observation. ***( P

    Article Snippet: Thereafter, anti-activated caspase3 antibody which was diluted 1:20 (Cell signaling, USA) was performed for overnight.

    Techniques: In Vivo, Mouse Assay, Staining

    IL-26 suppressed the apoptosis of HSCs by inhibition of caspase 3 (CASP3) and Bcl-2 associated X protein (BAX). A. Apoptosis of HSCs by annexin V-FITC/PI double staining after treatment with IL-26 at 0, 50, 100, and 200 pg/ml for 48 h. B. The mRNA expression levels of CASP3 and BAX were detected by quantitative real-time PCR (qRT-PCR) in the different concentrations of IL-26 groups. The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as the reference control. C and D. Western blotting analysis of the protein levels of CASP3, cleaved CASP3, and BAX in HSCs after IL-26 treatment. * P

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Interleukin-26 promotes the proliferation and activation of hepatic stellate cells to exacerbate liver fibrosis by the TGF-β1/Smad2 signaling pathway

    doi:

    Figure Lengend Snippet: IL-26 suppressed the apoptosis of HSCs by inhibition of caspase 3 (CASP3) and Bcl-2 associated X protein (BAX). A. Apoptosis of HSCs by annexin V-FITC/PI double staining after treatment with IL-26 at 0, 50, 100, and 200 pg/ml for 48 h. B. The mRNA expression levels of CASP3 and BAX were detected by quantitative real-time PCR (qRT-PCR) in the different concentrations of IL-26 groups. The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as the reference control. C and D. Western blotting analysis of the protein levels of CASP3, cleaved CASP3, and BAX in HSCs after IL-26 treatment. * P

    Article Snippet: Membranes were blocked with 5% non-fat milk at 37°C for 2 h and primary mouse monoclonal anti-TGF-β1 (#ab27969, 1:1000; Abcam, Cambridge, MA, USA) and anti-Bcl-2 associated X protein (BAX) (#ab77566, 1:500; Abcam) antibodies, and primary rabbit monoclonal anti-SMAD family member 2 (Smad2) (#ab40855, 1:500; Abcam), anti-caspase 3 (CASP3) (#9662, 1:1000; Cell Signaling Technology, Danvers, MA, USA), anti-cleaved CASP3 (#9661, 1:1500; Cell Signaling Technology) antibodies were prepared in blocking solution and incubated overnight at 4°C.

    Techniques: Inhibition, Double Staining, Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot

    EnvHA structures are defective in responses to growth regulatory signals. (A) Confocal cross sections through the center of acini stained with phalloidin (red) and immunostained with anti-Ki-67 (green) at 10 and 20 days in culture. Magnification, ×400; bar, 50 μm. (B) Confocal cross sections through the center of acini stained with DAPI (blue) and phalloidin (red) and immunostained with anti-c-caspase-3 (green) at 5 and 20 days in culture. Magnification, ×400; bar, 50 μm. *, c-caspase-3-positive cells. (C) Cell death in acini was monitored by ethidium bromide (EtBr) staining (1 μM) at 15 days of culture. Acini were analyzed by fluorescence microscopy; representative images are shown. Magnification, ×200; bar, 100 μm. (D) Confocal cross sections of acini that were immunostained with an antibody to ZO-1 (green) and counterstained with DAPI (blue) are depicted. Magnification, ×400; bars, 50 μm.

    Journal: Journal of Virology

    Article Title: Jaagsiekte Sheep Retrovirus Transformation in Madin-Darby Canine Kidney Epithelial Cell Three-Dimensional Culture ▿

    doi: 10.1128/JVI.02323-09

    Figure Lengend Snippet: EnvHA structures are defective in responses to growth regulatory signals. (A) Confocal cross sections through the center of acini stained with phalloidin (red) and immunostained with anti-Ki-67 (green) at 10 and 20 days in culture. Magnification, ×400; bar, 50 μm. (B) Confocal cross sections through the center of acini stained with DAPI (blue) and phalloidin (red) and immunostained with anti-c-caspase-3 (green) at 5 and 20 days in culture. Magnification, ×400; bar, 50 μm. *, c-caspase-3-positive cells. (C) Cell death in acini was monitored by ethidium bromide (EtBr) staining (1 μM) at 15 days of culture. Acini were analyzed by fluorescence microscopy; representative images are shown. Magnification, ×200; bar, 100 μm. (D) Confocal cross sections of acini that were immunostained with an antibody to ZO-1 (green) and counterstained with DAPI (blue) are depicted. Magnification, ×400; bars, 50 μm.

    Article Snippet: Antibodies used in analysis included Ki-67 (1:200; Zymed), c-caspase-3 (1:300; Cell Signaling), GM130 (1:300; BD Biosciences), HA (1:200; Cell Signaling), and ZO-1 (20 μg/ml; Invitrogen).

    Techniques: Staining, Fluorescence, Microscopy

    ( A ) Western immunoblot showing phospho-MAPK in Sca-1 + cells preconditioned with 100 nM IGF-1 for different time durations. Parallel experiment was performed with pre-treatment of Sca-1 + cells with 50 μM of MAPK blocker PD98059 for 30 min followed by 100 nM IGF-1 for the stipulated time durations. Western blot for p42/44 MAPK with/without PD98059 was performed on the same membrane and subjected to the same antibody incubation, detection, and exposure. The blots were later edited to align for comparison. ( B and C ) Densitometry for p42 MAPK and p44 MAPK for different durations of treatment showed the kinetics of MAPK phosphorylation over time and abrogation after PD98059 blocker treatment. ( D ) Representative western immunoblot from subfractionated cell lysate samples from preconditioning of Sca-1 + cells showing enhanced mitochondrial translocation of Cx-43 which was blocked by 50 μM PD98059 for 30 min (densitometric results are an average of three separate experiments). ( E ) Parallel experiments were performed to show that caspase-3 activity was significantly increased in the preconditioned Sca-1 + cells pre-treated with PD98059 with concomitant reduction in cell survival as determined by LDH release assay ( F ) (the number of experiments performed, n = 3).

    Journal: Cardiovascular Research

    Article Title: Mitochondria-specific transgenic overexpression of connexin-43 simulates preconditioning-induced cytoprotection of stem cells

    doi: 10.1093/cvr/cvq293

    Figure Lengend Snippet: ( A ) Western immunoblot showing phospho-MAPK in Sca-1 + cells preconditioned with 100 nM IGF-1 for different time durations. Parallel experiment was performed with pre-treatment of Sca-1 + cells with 50 μM of MAPK blocker PD98059 for 30 min followed by 100 nM IGF-1 for the stipulated time durations. Western blot for p42/44 MAPK with/without PD98059 was performed on the same membrane and subjected to the same antibody incubation, detection, and exposure. The blots were later edited to align for comparison. ( B and C ) Densitometry for p42 MAPK and p44 MAPK for different durations of treatment showed the kinetics of MAPK phosphorylation over time and abrogation after PD98059 blocker treatment. ( D ) Representative western immunoblot from subfractionated cell lysate samples from preconditioning of Sca-1 + cells showing enhanced mitochondrial translocation of Cx-43 which was blocked by 50 μM PD98059 for 30 min (densitometric results are an average of three separate experiments). ( E ) Parallel experiments were performed to show that caspase-3 activity was significantly increased in the preconditioned Sca-1 + cells pre-treated with PD98059 with concomitant reduction in cell survival as determined by LDH release assay ( F ) (the number of experiments performed, n = 3).

    Article Snippet: Sca-1+ cells were exposed to OGD, cell lysates were collected, and caspase-3 activity was measured with caspase-3 immunoassay kit as per instructions of the manufacturer (EMD Bioscience).

    Techniques: Western Blot, Incubation, Translocation Assay, Activity Assay, Lactate Dehydrogenase Assay

    C23 suppressed the transcriptional activity of p53 upon DNA damage and hypoxia A. pGL3-3×p53-BS-LUC and Renilla were co-transfected in HCT116 cells combined with and without C23 shRNA. 24 hours after transfection, cells were treated with Doxorubicin (100ng/ml) or upon 1% O 2 for indicated times and the cell lysates were analyzed by lucifease assay. B. HCT116 cells with and without stable overexpressing C23 were co-transfected with pGL3-3×p53-BS-LUC and Renilla. 24 hours after transfection, cells were treated with Doxorubicin (100ng/ml) or upon 1% O 2 for indicated times and then the cell lysates were subjected to lucifease assays. C. HCT116 cells with and without stable knockdown of C23 were treated with Doxorubicin (100ng/ml) for 24 hours, then p53 target gene mRNA expression levels were analyzed by qRT-PCR analysis. D-E. HCT116 cells with and without stable overexpressing C23 (a) or stable knockdown of C23(b) were treated with Doxorubicin (100ng/ml) or mock control for 24 hours. Cell lysates were then subjected to Western blot analysis with the indicated antibodies (D) and caspase3/7 activity analysis (E), respectively. F. (a) HCT116 cells stably expressing the control shRNA, C23 shRNA, p53 shRNA,or C23 shRNA plus p53 shRNA. (b) HCT116 cells with and without p53 knockdown were introduced with C23 respectively. Protein level of C23 and p53 were evaluated by Western blot. GAPDH served as loading control. G. HCT116 cells stably expressing the control shRNA, C23 shRNA, p53 shRNA,or C23 shRNA plus p53 shRNA (a) and HCT116 cells stably knocked down p53 were additionally introduced with C23 respectively (b). Cell number was evaluated by cell counter. The data were represented as mean±S.D. from three independent experiments. H. (a) HCT116 cells stably expressing the control shRNA, C23 shRNA, p53 shRNA,or C23 shRNA plus p53 shRNA. (b) HCT116 cells stably knocked down p53 were co-transfected with C23 cDNA respectively. Cells were then treated with 100ng/ml Doxorubicin for 24 hours. Cell viability was determined by MTT assay. The data were shown as the mean±s.d. of three independent experiments.

    Journal: Oncotarget

    Article Title: C23 promotes tumorigenesis via suppressing p53 activity

    doi: 10.18632/oncotarget.11071

    Figure Lengend Snippet: C23 suppressed the transcriptional activity of p53 upon DNA damage and hypoxia A. pGL3-3×p53-BS-LUC and Renilla were co-transfected in HCT116 cells combined with and without C23 shRNA. 24 hours after transfection, cells were treated with Doxorubicin (100ng/ml) or upon 1% O 2 for indicated times and the cell lysates were analyzed by lucifease assay. B. HCT116 cells with and without stable overexpressing C23 were co-transfected with pGL3-3×p53-BS-LUC and Renilla. 24 hours after transfection, cells were treated with Doxorubicin (100ng/ml) or upon 1% O 2 for indicated times and then the cell lysates were subjected to lucifease assays. C. HCT116 cells with and without stable knockdown of C23 were treated with Doxorubicin (100ng/ml) for 24 hours, then p53 target gene mRNA expression levels were analyzed by qRT-PCR analysis. D-E. HCT116 cells with and without stable overexpressing C23 (a) or stable knockdown of C23(b) were treated with Doxorubicin (100ng/ml) or mock control for 24 hours. Cell lysates were then subjected to Western blot analysis with the indicated antibodies (D) and caspase3/7 activity analysis (E), respectively. F. (a) HCT116 cells stably expressing the control shRNA, C23 shRNA, p53 shRNA,or C23 shRNA plus p53 shRNA. (b) HCT116 cells with and without p53 knockdown were introduced with C23 respectively. Protein level of C23 and p53 were evaluated by Western blot. GAPDH served as loading control. G. HCT116 cells stably expressing the control shRNA, C23 shRNA, p53 shRNA,or C23 shRNA plus p53 shRNA (a) and HCT116 cells stably knocked down p53 were additionally introduced with C23 respectively (b). Cell number was evaluated by cell counter. The data were represented as mean±S.D. from three independent experiments. H. (a) HCT116 cells stably expressing the control shRNA, C23 shRNA, p53 shRNA,or C23 shRNA plus p53 shRNA. (b) HCT116 cells stably knocked down p53 were co-transfected with C23 cDNA respectively. Cells were then treated with 100ng/ml Doxorubicin for 24 hours. Cell viability was determined by MTT assay. The data were shown as the mean±s.d. of three independent experiments.

    Article Snippet: The following antibodies were used in this study: GFP (Life, A11122 and Clonetech 632381), Flag (Sigma, F3165), GAPDH (Cell Signaling, 5174S), C23 (Santa Cruz Biotechnology, SC-8031 and Cell Signaling, 14574S), Caspase3 (Stressgene, AAP-113) and p53 (ABGENT, AJ1573a and Santa Cruz Biotechnology, SC-126).

    Techniques: Activity Assay, Transfection, shRNA, Expressing, Quantitative RT-PCR, Western Blot, Stable Transfection, MTT Assay

    Effect of almorexant and suvorexant on intracellular Ca 2+ release, cell growth and caspase-3 activity ( A ) Intracellular Ca 2+ production was detected in HEK-293 cells expressing recombinant native OX1R using Fluoforte Calcium Assay Kit (Enzo Life Sciences, NY, USA). Cells were challenged with 1 μM of OxA (top panel) or 1 μM OxA after preincubation with 10 μM suvorexant (middle panel) or 10 μM almorexant (bottom panel). ( B ) AsPC-1 cells were incubated for 48 h with OxA or almorexant or suvorexant in the presence (right panel) or in the absence (left panel) of 50 μM NSC87877. ( C ) colorometric Caspase-3 activity detection at 405 nm in AsPC-1 cells incubated with 1 μM OxA or 1 μM almorexant in the presence or in the absence of 50 μM NSC87877. * p

    Journal: Oncotarget

    Article Title: In vitro, in vivo and ex vivo demonstration of the antitumoral role of hypocretin-1/orexin-A and almorexant in pancreatic ductal adenocarcinoma

    doi: 10.18632/oncotarget.24084

    Figure Lengend Snippet: Effect of almorexant and suvorexant on intracellular Ca 2+ release, cell growth and caspase-3 activity ( A ) Intracellular Ca 2+ production was detected in HEK-293 cells expressing recombinant native OX1R using Fluoforte Calcium Assay Kit (Enzo Life Sciences, NY, USA). Cells were challenged with 1 μM of OxA (top panel) or 1 μM OxA after preincubation with 10 μM suvorexant (middle panel) or 10 μM almorexant (bottom panel). ( B ) AsPC-1 cells were incubated for 48 h with OxA or almorexant or suvorexant in the presence (right panel) or in the absence (left panel) of 50 μM NSC87877. ( C ) colorometric Caspase-3 activity detection at 405 nm in AsPC-1 cells incubated with 1 μM OxA or 1 μM almorexant in the presence or in the absence of 50 μM NSC87877. * p

    Article Snippet: Slides were immunolabeled with antibodies against OX1R (Life Technology, PA5-33837, polyclonal rabbit, 1/100), activated caspase-3 (Abgent, E87-77, polyclonal rabbit, 1/100) or Ki-67 (DAKO, clone MIB-1, monoclonal mouse, 1/100).

    Techniques: Activity Assay, Expressing, Recombinant, Calcium Assay, Incubation

    Effect of orexin-A on apoptosis in AsPC-1 cells - ( A ), SHP-2 protein tyrosine phosphatase inhibitor, NSC-87877, blocks orexin-induced apoptosis. AsPC-1 cells were challenged with (black bars) or without (white bars) 1 μM orexin-A for 48 hr in the absence or presence of NSC-87877 (50 μM). Apoptosis was measured by determination of annexin V-PE binding, and results are expressed as the percentage of apoptotic cells; ( B ) and ( C ). Indirect immunostaining of activated caspase-3 in AsPC-1 cells in the presence or absence of orexin-A. Paraformaldehyde-fixed AsPC-1 cells were challenged with (orexin-A) or without (basal) 1 μM orexin-A for 48 hr. Activated caspase-3 immunostaining is shown in B and scored in C. Results are means ± SE of three separate experiments. *** p

    Journal: Oncotarget

    Article Title: In vitro, in vivo and ex vivo demonstration of the antitumoral role of hypocretin-1/orexin-A and almorexant in pancreatic ductal adenocarcinoma

    doi: 10.18632/oncotarget.24084

    Figure Lengend Snippet: Effect of orexin-A on apoptosis in AsPC-1 cells - ( A ), SHP-2 protein tyrosine phosphatase inhibitor, NSC-87877, blocks orexin-induced apoptosis. AsPC-1 cells were challenged with (black bars) or without (white bars) 1 μM orexin-A for 48 hr in the absence or presence of NSC-87877 (50 μM). Apoptosis was measured by determination of annexin V-PE binding, and results are expressed as the percentage of apoptotic cells; ( B ) and ( C ). Indirect immunostaining of activated caspase-3 in AsPC-1 cells in the presence or absence of orexin-A. Paraformaldehyde-fixed AsPC-1 cells were challenged with (orexin-A) or without (basal) 1 μM orexin-A for 48 hr. Activated caspase-3 immunostaining is shown in B and scored in C. Results are means ± SE of three separate experiments. *** p

    Article Snippet: Slides were immunolabeled with antibodies against OX1R (Life Technology, PA5-33837, polyclonal rabbit, 1/100), activated caspase-3 (Abgent, E87-77, polyclonal rabbit, 1/100) or Ki-67 (DAKO, clone MIB-1, monoclonal mouse, 1/100).

    Techniques: Binding Assay, Immunostaining

    ( A ) The effects on Bax, Bcl-2, caspase-3, caspase-9, TNF-α, JNK, and p-JNK protein expression levels were detected by the Western blot method after exposure to MMO for 24 h; ( B ) Protein expression level of bax/bcl-2 was detected by Western blot after exposure to MMO; ( C ) Protein expression level of Caspase-3/β-actin was detected by Western blot after exposure to MMO; ( D ) Protein expression level of Caspase-9/β-actin was detected by Western blot after exposure to MMO; ( E ) Protein expression level of TNF-α/β-actin was detected by Western blot after exposure to MMO; ( F ) Protein expression level of p-JNK/β-actin was detected by Western blot after exposure to MMO; ( G ) Protein expression level of JNK/β-actin was detected by Western blot after exposure to MMO. β-actin was used as a control in all histograms; ( H ) SDS-PAGE image stained with Coomassie brilliant blue. Data are presented as mean ± S.D. for three independent experiments with triplicate determination.* p

    Journal: Marine Drugs

    Article Title: Protective Effects and Mechanism of Meretrix meretrix Oligopeptides against Nonalcoholic Fatty Liver Disease

    doi: 10.3390/md15020031

    Figure Lengend Snippet: ( A ) The effects on Bax, Bcl-2, caspase-3, caspase-9, TNF-α, JNK, and p-JNK protein expression levels were detected by the Western blot method after exposure to MMO for 24 h; ( B ) Protein expression level of bax/bcl-2 was detected by Western blot after exposure to MMO; ( C ) Protein expression level of Caspase-3/β-actin was detected by Western blot after exposure to MMO; ( D ) Protein expression level of Caspase-9/β-actin was detected by Western blot after exposure to MMO; ( E ) Protein expression level of TNF-α/β-actin was detected by Western blot after exposure to MMO; ( F ) Protein expression level of p-JNK/β-actin was detected by Western blot after exposure to MMO; ( G ) Protein expression level of JNK/β-actin was detected by Western blot after exposure to MMO. β-actin was used as a control in all histograms; ( H ) SDS-PAGE image stained with Coomassie brilliant blue. Data are presented as mean ± S.D. for three independent experiments with triplicate determination.* p

    Article Snippet: Antibodies against Caspase-3, TNF-α, P-JNK, and JNK1/2/3 were purchased from ABGENT (San Diego, CA, USA).

    Techniques: Expressing, Western Blot, SDS Page, Staining

    Heat stress induced apoptosis and caspase-3 activation. F98 cells were incubated at 43°C for 60 min to simulate heat stress. Culture medium was then replaced prior to further incubation of the cells for 0, 3, 6 or 12 h (R0 to R12, respectively). (A) Analysis of apoptosis was performed with flow cytometry using Annexin V-FITC/PI staining. (B) The fluorogenic substrate Ac-DEVD-AMC was applied to measure enzymatic activity of caspase-3, which was expressed relative to the control cells incubated at 37°C (set as 100%). *P

    Journal: Oncology Letters

    Article Title: p38 MAPK-MK2 pathway regulates the heat-stress-induced accumulation of reactive oxygen species that mediates apoptotic cell death in glial cells

    doi: 10.3892/ol.2017.7360

    Figure Lengend Snippet: Heat stress induced apoptosis and caspase-3 activation. F98 cells were incubated at 43°C for 60 min to simulate heat stress. Culture medium was then replaced prior to further incubation of the cells for 0, 3, 6 or 12 h (R0 to R12, respectively). (A) Analysis of apoptosis was performed with flow cytometry using Annexin V-FITC/PI staining. (B) The fluorogenic substrate Ac-DEVD-AMC was applied to measure enzymatic activity of caspase-3, which was expressed relative to the control cells incubated at 37°C (set as 100%). *P

    Article Snippet: The following rabbit primary antibodies were used at a 1:2,000 dilution: GAPDH (cat. no. ab70699; Abcam), phosphorylated (p)-JNK (cat. no. 4668; Cell Signaling Technology, Inc., Danvers, MA, USA), p-p38 (cat. no. 4511; Cell Signaling Technology, Inc.), p-ERK1/2 (cat. no. 4370; Cell Signaling Technology, Inc.), JNK (cat. no. 9252; Cell Signaling Technology, Inc.), p38 (cat. no. 8690; Cell Signaling Technology, Inc.), ERK1/2 (cat. no. 4695; Cell Signaling Technology, Inc.), MK2 (cat. no. ab131531; Abcam), p-MK2 (cat. no. ab63378; Abcam), MK5 (cat. no. 7419; Cell Signaling Technology, Inc.), p-MK5 (cat. no. 9771; Cell Signaling Technology, Inc.), caspase-3 (cat. no. 14220S; Cell Signaling Technology, Inc.) and cleaved caspase-3 (cat. no. 9654S; Cell Signaling Technology, Inc.) overnight at 4°C.

    Techniques: Activation Assay, Incubation, Flow Cytometry, Cytometry, Staining, Activity Assay

    Effect of cell density over IDS-ko NSCs differentiation. ( a–f ) Coimmunostaining for caspase3+ and ubiquitin with the neural differentiation markers β -tubIII, GFAP and GalC of wt and IDS-ko NSCs at 0 and 17 div of differentiation under HD (1.25 × 10 4 cells/cm 2 ) and LD (6.5 × 10 3 cells/cm 2 ) cell density conditions. ( a ) Confocal microscopy images showing the coimmunostaining of caspase3 marker with GFAP, GalC or β -tubIII under HD or low LD conditions. Scale bar: 100 μ m. ( b ) Quantification of GFAP+, β -tubIII+ and GalC+ cells over total DAPI+ nuclei under HD and LD conditions. ( c ) Quantification of caspase3+ cells over total DAPI+ nuclei under HD and LD conditions. ( d ) Quantification of GFAP+/caspase3+, β -tubIII/caspase3+, GalC+/caspase3+ cells over total caspase3+ cells under HD and LD conditions. ( e) Confocal microscopy images showing the immunostaining of ubiquitin+ marker. Scale bar: 100 μ m. ( f ) Quantification of ubiquitin+ cells over total DAPI+ nuclei under HD and LD conditions. ( g ) Confocal microscopy images showing the presence of caspase3+ cells in wt (upper left) and IDS-ko (upper right) mouse cortex. Scale bar: 75 μ m; in inset: 11.72 μ m. Quantification of caspase3+ cells over total DAPI+ nuclei (bottom left) and colocalization with glial markers GFAP (bottom center) and MBP (bottom right). Scale bars: 21, 10 μ m. ( h ) Confocal microscopy images showing the presence of ubiquitin+ cells in wt (upper left) and IDS-ko (upper right) mouse cortex. Scale bar: 75 μ m. Inset scale bar: 16 μ m. Quantification of ubiquitin+ cells over total DAPI+ nuclei (bottom left) and colocalization with GFAP (bottom center) and MBP (bottom right). Scale bar: 13.7 μ m. ( i ) Confocal microscopy images showing the presence of caspase3+ cells in wt (upper left) and Hunter (upper right) human cortex (end stage). Scale bar: 75 μ m. Quantification of caspase3+ cells over total DAPI+ nuclei (bottom left) and colocalization with GFAP (bottom center), MBP (bottom right) and neuronal marker β -tubIII (bottom right). Scale bar: 10–14 μ m. ( j) Confocal microscopy images showing the presence of ubiquitin+ aggregates in wt (upper left) and Hunter (upper right) human cortex (end stage). Scale bar: 75 μ m. Quantification of caspase3+ cells over total DAPI+ nuclei (bottom left) and colocalization with GFAP (bottom center), MBP (bottom right) and β -tubIII (bottom right). Scale bars: 16–18 μ m. Student's t -test was applied, the differences among all the values were statistically not significant unless indicated (* P ≤0.05,** P ≤0.01, *** P ≤0.001) values are means±S.E.M. Casp, active caspase 3; Ub, ubiquitin

    Journal: Cell Death & Disease

    Article Title: Murine neural stem cells model Hunter disease in vitro: glial cell-mediated neurodegeneration as a possible mechanism involved

    doi: 10.1038/cddis.2013.430

    Figure Lengend Snippet: Effect of cell density over IDS-ko NSCs differentiation. ( a–f ) Coimmunostaining for caspase3+ and ubiquitin with the neural differentiation markers β -tubIII, GFAP and GalC of wt and IDS-ko NSCs at 0 and 17 div of differentiation under HD (1.25 × 10 4 cells/cm 2 ) and LD (6.5 × 10 3 cells/cm 2 ) cell density conditions. ( a ) Confocal microscopy images showing the coimmunostaining of caspase3 marker with GFAP, GalC or β -tubIII under HD or low LD conditions. Scale bar: 100 μ m. ( b ) Quantification of GFAP+, β -tubIII+ and GalC+ cells over total DAPI+ nuclei under HD and LD conditions. ( c ) Quantification of caspase3+ cells over total DAPI+ nuclei under HD and LD conditions. ( d ) Quantification of GFAP+/caspase3+, β -tubIII/caspase3+, GalC+/caspase3+ cells over total caspase3+ cells under HD and LD conditions. ( e) Confocal microscopy images showing the immunostaining of ubiquitin+ marker. Scale bar: 100 μ m. ( f ) Quantification of ubiquitin+ cells over total DAPI+ nuclei under HD and LD conditions. ( g ) Confocal microscopy images showing the presence of caspase3+ cells in wt (upper left) and IDS-ko (upper right) mouse cortex. Scale bar: 75 μ m; in inset: 11.72 μ m. Quantification of caspase3+ cells over total DAPI+ nuclei (bottom left) and colocalization with glial markers GFAP (bottom center) and MBP (bottom right). Scale bars: 21, 10 μ m. ( h ) Confocal microscopy images showing the presence of ubiquitin+ cells in wt (upper left) and IDS-ko (upper right) mouse cortex. Scale bar: 75 μ m. Inset scale bar: 16 μ m. Quantification of ubiquitin+ cells over total DAPI+ nuclei (bottom left) and colocalization with GFAP (bottom center) and MBP (bottom right). Scale bar: 13.7 μ m. ( i ) Confocal microscopy images showing the presence of caspase3+ cells in wt (upper left) and Hunter (upper right) human cortex (end stage). Scale bar: 75 μ m. Quantification of caspase3+ cells over total DAPI+ nuclei (bottom left) and colocalization with GFAP (bottom center), MBP (bottom right) and neuronal marker β -tubIII (bottom right). Scale bar: 10–14 μ m. ( j) Confocal microscopy images showing the presence of ubiquitin+ aggregates in wt (upper left) and Hunter (upper right) human cortex (end stage). Scale bar: 75 μ m. Quantification of caspase3+ cells over total DAPI+ nuclei (bottom left) and colocalization with GFAP (bottom center), MBP (bottom right) and β -tubIII (bottom right). Scale bars: 16–18 μ m. Student's t -test was applied, the differences among all the values were statistically not significant unless indicated (* P ≤0.05,** P ≤0.01, *** P ≤0.001) values are means±S.E.M. Casp, active caspase 3; Ub, ubiquitin

    Article Snippet: After blocking with 10% normal goat serum, cultures were incubated overnight at 4 °C with the following antibodies (monoclonal antibody, monoclonal; polyclonal antibody, polyclonal): nestin (monoclonal, MAB 353, Millipore, Bedford, MA, USA; 1:200, Species Cross Reactivity: mouse, rat), β -tubIII (monoclonal, MMS-435P, Covance, Princeton, NJ, USA; 1:400, Species Cross Reactivity: mammalian), GFAP (polyclonal, Dako, Glostrup, Denmark; 1:400, Species Cross Reactivity: mammalian), GalC (monoclonal, MAB345, Chemicon - Millipore Bioscience Research Reagents, Temecula, CA, USA; 1:100, Species Cross Reactivity: bovine, human, mouse, rat, rabbit), Lamp1 (polyclonal, ab24170, Abcam, Cambridge, UK; 1:750, Species Cross Reactivity: dog, human, mouse, rat, Zebrafish), caspase3 (Asp 170) (polyclonal, #9961, Cell Signaling, Danvers, MA, USA; 1:500, Species Cross Reactivity: human, mouse, rat, monkey, bovine), ubiquitin (polyclonal, Z0458, Dako, 1:50, Species Cross Reactivity: human, mouse, rat), MAP2 (monoclonal, Chemicon, 1:400, Species Cross Reactivity: human, rat, mouse, bovine, chicken), KI67 nuclear antigen (polyclonal, Novocastra - Leica Microsystems GmbH, Wetzlar, Germany; NCL-Ki67p, 1:1000, Species Cross Reactivity: human, mouse), PDGFRα (PDGFRα , polyclonal, #3164, Cell Signaling, 1:100, Species Cross Reactivity: human, mouse, rat).

    Techniques: Confocal Microscopy, Marker, Immunostaining

    Placement of the VOC, umbilical artery, and OUA connection is dose-dependent on T (A) Dot-box plot of CASP3+ cells in allantois and overlying yolk sac (EB-6s stages); means (dashed line), SEM (error bars); n.s., not significant (Student t -Test; T + /T + vs T C /T + , P =0.8912; T + /T + vs T C /T C , P =0.3714; T C /T + vs T C /T C , P =0.2134). ( B-D ) Immunostaining for FLK-1 amongst T C genotypes. Black boxed insets, lower left: enlarged prospective VOC of black-boxed region in main panels. Black arrows (main panels) and arrowheads (insets): correctly patterned FLK-1 angioblasts within distal allantois and prospective VOC. Dashed lines, posteriormost extension of primitive streak (B-D); white asterisk, ACD (B, C). Magenta inset (D): a misplaced PECAM-1-positive (magenta arrow) vessel within the yolk sac near its junction with the allantois, the region of which is indicated by the magenta box in the main panel (see text). ( E, F ) 3D models (frontal ventral views) reconstructed from PECAM-1-immunostained nascent arterial vessels at the posterior embryonic-extraembryonic interface; dashed vertical line (E, F) marks the axial midline. Color key: blue, dorsal aortae (da); red, VOC (asterisk); yellow, umbilical artery (ua). ( G ) T C /T C mutant specimen immunostained for PECAM-1, sagittal section, lateral to the midline, equivalent to the solid horizontal line in F. White arrow indicates a vessel forming at the allantoic-yolk sac junction off the midline. ( H-K ) Examples of tissue sections used for the 3D reconstructions in (L-N). (H) PECAM-1-immunostained transverse section showing the paired da on either side of the hindgut (hg), and the omphalomesenteric artery (oa) at the ventral midline. (I-K) Colorized transverse sections from the level of the oa (H, I) and proceeding posteriorly (distally), to capture the VOC (J) and ua (K). All reconstructed sections were PECAM-1 immunostained. ( L-N ) Top, ventral (frontal) and side views (posterior, right) of the OUA connection in all three T genotypes, colored as described above with addition of purple for the oa. Red arrows (N) indicate missing OUA connection; vertical dashed line indicates the axial midline and the site where the VOC (red, located off center) should be located. Scale bar (G): 10 μm (D magenta inset); 20 μm (B-D, H-K), 25 μm (G). al, allantois; am, amnion; em, embryo; ys, yolk sac.

    Journal: Developmental biology

    Article Title: Brachyury drives formation of a distinct vascular branchpoint critical for fetal-placental arterial union in the mouse gastrula

    doi: 10.1016/j.ydbio.2017.03.032

    Figure Lengend Snippet: Placement of the VOC, umbilical artery, and OUA connection is dose-dependent on T (A) Dot-box plot of CASP3+ cells in allantois and overlying yolk sac (EB-6s stages); means (dashed line), SEM (error bars); n.s., not significant (Student t -Test; T + /T + vs T C /T + , P =0.8912; T + /T + vs T C /T C , P =0.3714; T C /T + vs T C /T C , P =0.2134). ( B-D ) Immunostaining for FLK-1 amongst T C genotypes. Black boxed insets, lower left: enlarged prospective VOC of black-boxed region in main panels. Black arrows (main panels) and arrowheads (insets): correctly patterned FLK-1 angioblasts within distal allantois and prospective VOC. Dashed lines, posteriormost extension of primitive streak (B-D); white asterisk, ACD (B, C). Magenta inset (D): a misplaced PECAM-1-positive (magenta arrow) vessel within the yolk sac near its junction with the allantois, the region of which is indicated by the magenta box in the main panel (see text). ( E, F ) 3D models (frontal ventral views) reconstructed from PECAM-1-immunostained nascent arterial vessels at the posterior embryonic-extraembryonic interface; dashed vertical line (E, F) marks the axial midline. Color key: blue, dorsal aortae (da); red, VOC (asterisk); yellow, umbilical artery (ua). ( G ) T C /T C mutant specimen immunostained for PECAM-1, sagittal section, lateral to the midline, equivalent to the solid horizontal line in F. White arrow indicates a vessel forming at the allantoic-yolk sac junction off the midline. ( H-K ) Examples of tissue sections used for the 3D reconstructions in (L-N). (H) PECAM-1-immunostained transverse section showing the paired da on either side of the hindgut (hg), and the omphalomesenteric artery (oa) at the ventral midline. (I-K) Colorized transverse sections from the level of the oa (H, I) and proceeding posteriorly (distally), to capture the VOC (J) and ua (K). All reconstructed sections were PECAM-1 immunostained. ( L-N ) Top, ventral (frontal) and side views (posterior, right) of the OUA connection in all three T genotypes, colored as described above with addition of purple for the oa. Red arrows (N) indicate missing OUA connection; vertical dashed line indicates the axial midline and the site where the VOC (red, located off center) should be located. Scale bar (G): 10 μm (D magenta inset); 20 μm (B-D, H-K), 25 μm (G). al, allantois; am, amnion; em, embryo; ys, yolk sac.

    Article Snippet: Primary antibodies, sources, stock concentrations, dilutions, and Research Resource Identifiers (RRIDs) were: Bi- Phospho-FGFR Y653/654 (pFGFR1) (AP3104A, Abgent, San Diego, CA; 0.25 mg/ml, rabbit polyclonal; 1/60 dilution; RRID: AB_ 636986); BRACHYURY (T) (sc-17743, Santa Cruz Biotechnologies, Santa Cruz, California; 200 μg/ml, goat polyclonal; 1/100 dilution; RRID: AB_634980); CASPASE-3 (CASP3) (559565, BD Biosciences, San Jose, California; 0.5 mg/ml, rabbit monoclonal; 1/100 dilution; RRID: AB_397274); FGF2 (sc-79-G, Santa Cruz Biotechnologies; 200 μg/ml, goat polyclonal; 1/50 dilution; RRID: AB_631498); FGFR1 (ab10646, Abcam, Cambridge, MA; 1500 μg/ml, rabbit polyclonal; 1/400 dilution; RRID: AB_297367); FGFR2 (sc-122-G, Santa Cruz Biotechnologies; 200 μg/ml, goat polyclonal; 1/300 dilution; RRID: AB_631510); FGFR3 (sc-123-G, Santa Cruz Biotechnologies; 200 μg/ml, goat polyclonal; 1/300 dilution); FLK-1 (sc-315-G, Santa Cruz Biotechnologies; 200 μg/ml, goat polyclonal; 1/50 dilution; RRID: AB_632599) ( ); OCT-3/4 (sc-8628, Santa Cruz Biotechnologies; 200 μg/ml, goat polyclonal; 1/100 dilution; RRID: AB_653551); and PECAM-1 (AF3628, R & D Systems, Minneapolis, MN; 0.2 mg/ml, goat polyclonal; 1/500 dilution; RRID: AB_2161028).

    Techniques: Immunostaining, Mutagenesis

    OLE affects Bcl-2 family, cleaved caspase-3 protein expression, and Caspase-3 activity on day 5 after IRI or sham. Representative Western blots and densitometric quantification of Bax/Bcl-2 ( A ). Representative Western blots and densitometric quantification and cleaved caspase-3/β-actin on day 5 after IRI ( B ). Caspase-3 activity in brain tissues of rats in different groups ( C ). Data are presented as the mean ±SEM (* P

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: Protective Effects of Oleuropein Against Cerebral Ischemia/Reperfusion by Inhibiting Neuronal Apoptosis

    doi: 10.12659/MSM.912336

    Figure Lengend Snippet: OLE affects Bcl-2 family, cleaved caspase-3 protein expression, and Caspase-3 activity on day 5 after IRI or sham. Representative Western blots and densitometric quantification of Bax/Bcl-2 ( A ). Representative Western blots and densitometric quantification and cleaved caspase-3/β-actin on day 5 after IRI ( B ). Caspase-3 activity in brain tissues of rats in different groups ( C ). Data are presented as the mean ±SEM (* P

    Article Snippet: After blocking with 5% fat-free dry milk for 2 h at room temperature, the membrane was then incubated overnight at 4˚C with primary antibody, including Akt (anti-rabbit, 1: 1000, Abcam), p-Akt (anti-rabbit, 1: 1000, Abcam), GSK3β (anti-rabbit, 1: 1000, Abcam), p-GSK3β (anti-rabbit, 1: 1000, Abcam), β-actin (anti-rabbit, 1: 1000, Abcam), cleaved caspase-3 (CC3, anti-mouse, 1: 1000; Cell Signaling), caspase-3 (anti-rabbit, 1: 1000, Abcam), B-cell leukemia/lymphoma 2 (Bcl-2, anti-rabbit, 1: 1000, Abcam), BCL2-Associated X (Bax, anti-rabbit, 1: 1000, Abcam), brain-derived neurotrophic factor (BDNF; anti-rabbit, 1: 1000, Abcam), and nerve growth factor (NGF; anti-rabbit, 1: 1000, Abcam).

    Techniques: Expressing, Activity Assay, Western Blot

    Upregulation of miR-34a-induced apoptosis of hepatocytes. Human normal hepatocytes cell line L-02 cells were transfected with miR-34a mimics (20, 50 nM) or negative miRNA control (NC) with or without SRT1720 for 48 h. Expression of miR-34a was confirmed (A) at 24 h after transfection. Changes in the expression of SIRT1, p53 and Ac-p53 were determined (B) after 48 h. The apoptosis of hepatocytes was detected using an Annexin V-FITC/PI Apoptosis Detection Kit in a flow cytometry system (C) or by detecting the cleavage of caspase3 (D). Blank, L-02 cells; NC, negative control (L-02 cells transfected with negative mimics). * P

    Journal: PLoS ONE

    Article Title: Activation of the miR-34a/SIRT1/p53 Signaling Pathway Contributes to the Progress of Liver Fibrosis via Inducing Apoptosis in Hepatocytes but Not in HSCs

    doi: 10.1371/journal.pone.0158657

    Figure Lengend Snippet: Upregulation of miR-34a-induced apoptosis of hepatocytes. Human normal hepatocytes cell line L-02 cells were transfected with miR-34a mimics (20, 50 nM) or negative miRNA control (NC) with or without SRT1720 for 48 h. Expression of miR-34a was confirmed (A) at 24 h after transfection. Changes in the expression of SIRT1, p53 and Ac-p53 were determined (B) after 48 h. The apoptosis of hepatocytes was detected using an Annexin V-FITC/PI Apoptosis Detection Kit in a flow cytometry system (C) or by detecting the cleavage of caspase3 (D). Blank, L-02 cells; NC, negative control (L-02 cells transfected with negative mimics). * P

    Article Snippet: The membranes were blocked with Tris-buffered saline (TBS) containing 0.1% Tween-20 (TBST) and 5% (w/v) nonfat dry milk for 1 h. The membranes were then incubated overnight at 4°C with the primary antibodies of α-smooth muscle actin (SMA), SIRT1, p53, acetylated p53 (Ac-p53), collagen I, caspase3, and β-actin (Abcam, Cambridge, MA) at a dilution of 1:1000.

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

    Mitochondrial fission factor (Mff) induced cardiac microcirculation endothelial cells (CMECs) apoptosis via excessive mitochondrial fission in vitro. Wild‐type (WT) mice– and homozygous Mff–deficient (Mff gt ) mice–derived CMECs were named WT and Mff gt groups, respectively. Furthermore, Mff gain‐of‐function experiments were performed in CMEC s from Mff gt using adenovirus vector (Ad+Mff gt group). Meanwhile, Mdivi‐1, an inhibitor of fission, was used in CMEC s from WT mice as the negative control group. The ischemia/reperfusion (IR) injury in vitro was mimicked by 30 minutes of hypoxia with serum starvation and 2 hours of reoxygeneration. A and B, Mitochondria of CMECs are labeled with anti‐Tom20 antibody to determine the number of cells with mitochondria fragmentation. The boxed area under each micrograph is enlarged to determine mitochondria fragmentation. To assess changes in mitochondrial morphology quantitatively, the aspect ratio ( AR ; the mitochondrial length) and form factor ( FF ; the degree of mitochondrial branching) were calculated for each cell (the minimum value for both parameters is 1). High FF and AR values show healthy mitochondria, whereas low FF and AR indicate fragmented mitochondria. C, Co‐localization of dynamin‐related protein 1 (Drp1) and mitochondria. The boxed area under each micrograph represents the amplification of the white square. More Drp1 was located on fragmented mitochondria while loss of Mff could reduce Drp1 migration on mitochondria. D through F, IR increased mitochondria‐Drp1 expression. Meanwhile, IR also reduced proteins related to mitochondrial fusion. The control of cytoplasm and mitochondrial fractionation in the Western blots are β‐actin and cytochrome c oxidase subunit IV , respectively. D and G, Caspase‐3 activation ( CL .caspase3 expression) was detected by Western blots. H, Mff and CL .caspase3 co‐location by immunofluorescence. I, Transendothelial electrical resistance ( TER ) and permeability examination in CMEC s subjected to IR injury. Confluence of CMEC s monolayer was assessed as stabilized basal resistance of 800 Ω. TER increases when endothelial cells adhere and spread out, and decreases when endothelial cells retract or lose adhesion, which is the marker of endothelial barrier function. J, Fluorescein isothiocyanate (FITC)–dextran clearance was measured to assess changes in endothelial permeability. FITC ‐dextran was added on top of the inserts, allowing it to permeate through the cell monolayer. The increased endothelial permeability could retain more FITC ‐dextran. Thus, the FITC content remaining on the plate after IR injury indicated the extent of permeability of CMEC s. * P

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Mff‐Dependent Mitochondrial Fission Contributes to the Pathogenesis of Cardiac Microvasculature Ischemia/Reperfusion Injury via Induction of mROS‐Mediated Cardiolipin Oxidation and HK2/ VDAC1 Disassociation‐Involved mPTP Opening

    doi: 10.1161/JAHA.116.005328

    Figure Lengend Snippet: Mitochondrial fission factor (Mff) induced cardiac microcirculation endothelial cells (CMECs) apoptosis via excessive mitochondrial fission in vitro. Wild‐type (WT) mice– and homozygous Mff–deficient (Mff gt ) mice–derived CMECs were named WT and Mff gt groups, respectively. Furthermore, Mff gain‐of‐function experiments were performed in CMEC s from Mff gt using adenovirus vector (Ad+Mff gt group). Meanwhile, Mdivi‐1, an inhibitor of fission, was used in CMEC s from WT mice as the negative control group. The ischemia/reperfusion (IR) injury in vitro was mimicked by 30 minutes of hypoxia with serum starvation and 2 hours of reoxygeneration. A and B, Mitochondria of CMECs are labeled with anti‐Tom20 antibody to determine the number of cells with mitochondria fragmentation. The boxed area under each micrograph is enlarged to determine mitochondria fragmentation. To assess changes in mitochondrial morphology quantitatively, the aspect ratio ( AR ; the mitochondrial length) and form factor ( FF ; the degree of mitochondrial branching) were calculated for each cell (the minimum value for both parameters is 1). High FF and AR values show healthy mitochondria, whereas low FF and AR indicate fragmented mitochondria. C, Co‐localization of dynamin‐related protein 1 (Drp1) and mitochondria. The boxed area under each micrograph represents the amplification of the white square. More Drp1 was located on fragmented mitochondria while loss of Mff could reduce Drp1 migration on mitochondria. D through F, IR increased mitochondria‐Drp1 expression. Meanwhile, IR also reduced proteins related to mitochondrial fusion. The control of cytoplasm and mitochondrial fractionation in the Western blots are β‐actin and cytochrome c oxidase subunit IV , respectively. D and G, Caspase‐3 activation ( CL .caspase3 expression) was detected by Western blots. H, Mff and CL .caspase3 co‐location by immunofluorescence. I, Transendothelial electrical resistance ( TER ) and permeability examination in CMEC s subjected to IR injury. Confluence of CMEC s monolayer was assessed as stabilized basal resistance of 800 Ω. TER increases when endothelial cells adhere and spread out, and decreases when endothelial cells retract or lose adhesion, which is the marker of endothelial barrier function. J, Fluorescein isothiocyanate (FITC)–dextran clearance was measured to assess changes in endothelial permeability. FITC ‐dextran was added on top of the inserts, allowing it to permeate through the cell monolayer. The increased endothelial permeability could retain more FITC ‐dextran. Thus, the FITC content remaining on the plate after IR injury indicated the extent of permeability of CMEC s. * P

    Article Snippet: The primary antibodies were as follows: CD31 (1:1500, Abcam), vascular endothelial cadherin (VE‐cadherin; 1:1000, Abcam), cyt‐c (1:500, Abcam), Mff (1:1000, Abcam), pDrp1 (1:500, Abcam), HK2 (1:500, Abcam), cleaved caspase3 (CL.caspase3; 1:1000, Abcam), and pro‐caspase3 (1:2000, Abcam).

    Techniques: In Vitro, Mouse Assay, Derivative Assay, Plasmid Preparation, Negative Control, Labeling, Amplification, Migration, Expressing, Fractionation, Western Blot, Activation Assay, Immunofluorescence, Permeability, Marker

    CQ protects embryonic brains from ZIKV infection and microcephaly. Littermate embryonic brains were injected with ZIKV or medium at E13.5, then the pregnant mice were treated daily with CQ or vehicle until E18.5. (A) Images of brains and Nissl staining of coronal sections. (B) Measurements of cortical layer thickness. Mock + Veh: n = 9/4, Mock + CQ: n = 6/3, ZIKV + Veh: n = 6/3, ZIKV + CQ: n = 8/4. CP: cortical plate, SP: subplate, IZ: intermediate zone, SVZ: subventricular zone, VZ: ventricular zone. (C) Images of coronal sections stained with ZIKV antiserum (green) and DAPI (blue). Right panel: Quantification of relative intensity. n = 7/3 for each. (D) Images of cortices stained for the activated form of Caspase3 (white) and DAPI (blue). Right panel: quantification of relative intensity. N = 7/3 for each. (E) Images of cortices stained with phosphorylated Histone H3 (P–H3, red). Right panel: quantification of P-H3 + cells. Mock + Veh: n = 7/4, Mock + CQ: n = 8/3, ZIKV + Veh: n = 5/3, ZIKV + CQ: n = 5/3. (F) Coronal sections stained for ZIKV (green), Sox2 (red) and Tbr2 (white). Right panel: quantification of the density of Sox2 + cells and Tbr2 + cells in the cortices. Mock + Veh: n = 8/4 (Sox2 +), 9/4 (Tbr2 +); Mock + CQ: n = 6/3 (Sox2 +), 6/3 (Tbr2 +); ZIKV + Veh: n = 10/5 (Sox2 +), 10/5 (Tbr2 +); ZIKV + CQ: n = 10/5 (Sox2 +), 10/5 (Tbr2 +). All data are means ± SEM. Student's t-test. * P

    Journal: EBioMedicine

    Article Title: Chloroquine, a FDA-approved Drug, Prevents Zika Virus Infection and its Associated Congenital Microcephaly in Mice

    doi: 10.1016/j.ebiom.2017.09.034

    Figure Lengend Snippet: CQ protects embryonic brains from ZIKV infection and microcephaly. Littermate embryonic brains were injected with ZIKV or medium at E13.5, then the pregnant mice were treated daily with CQ or vehicle until E18.5. (A) Images of brains and Nissl staining of coronal sections. (B) Measurements of cortical layer thickness. Mock + Veh: n = 9/4, Mock + CQ: n = 6/3, ZIKV + Veh: n = 6/3, ZIKV + CQ: n = 8/4. CP: cortical plate, SP: subplate, IZ: intermediate zone, SVZ: subventricular zone, VZ: ventricular zone. (C) Images of coronal sections stained with ZIKV antiserum (green) and DAPI (blue). Right panel: Quantification of relative intensity. n = 7/3 for each. (D) Images of cortices stained for the activated form of Caspase3 (white) and DAPI (blue). Right panel: quantification of relative intensity. N = 7/3 for each. (E) Images of cortices stained with phosphorylated Histone H3 (P–H3, red). Right panel: quantification of P-H3 + cells. Mock + Veh: n = 7/4, Mock + CQ: n = 8/3, ZIKV + Veh: n = 5/3, ZIKV + CQ: n = 5/3. (F) Coronal sections stained for ZIKV (green), Sox2 (red) and Tbr2 (white). Right panel: quantification of the density of Sox2 + cells and Tbr2 + cells in the cortices. Mock + Veh: n = 8/4 (Sox2 +), 9/4 (Tbr2 +); Mock + CQ: n = 6/3 (Sox2 +), 6/3 (Tbr2 +); ZIKV + Veh: n = 10/5 (Sox2 +), 10/5 (Tbr2 +); ZIKV + CQ: n = 10/5 (Sox2 +), 10/5 (Tbr2 +). All data are means ± SEM. Student's t-test. * P

    Article Snippet: The antibodies used for immunostaining were anti-ZIKV serum from a patient (1:1000), Activated-caspase3 (Abcam, ab2302, 1:1000), Phospho-Histone 3 (P-H3) (Abcam, ab10543, 1:1000), Sox2 (Abcam, ab97959, 1:1000), Tbr2 (Miilipore, ab15894, 1:1000), NeuN (Abcam, ab104224, 1:1000), and Tbr1 (Abcam, ab31940, 1:500).

    Techniques: Infection, Injection, Mouse Assay, Staining

    PLY induces endoplasmic reticulum (ER) stress pathway without progressing to apoptosis in HL-1 cardiomyocytes. ( A ) Typical Western blots showing the activation of ER stress markers, (p-elF2α, p-IRE, BiP), JNK and ERK in HL-1 cells 30 min after PLY treatment. ( B ) Representative western blots showing the effect of PLY on the induction of apoptotic markers CHOP and active caspase-3 after 8 hour of treatment. Thapsigargin (Tg 5 μM) and Staurosporin (Stau 10 μM) were used as positive inducers of CHOP and active caspase-3 respectively. ( C-G ) Effects of PLY (1.0 μg/ml) and PLY+ 4-Phenylbutyric acid (4-PBA, an ER stress inhibitor, 10 mM) on Peak Shortening (C), (+dL/dt) (D), TTP (E), tR 90 (F) and (-dL/dt) (G) of HL-1 cells after 30 min treatment are presented as Mean±SD (n = 9 from 3 independent experiments). * p

    Journal: PLoS Pathogens

    Article Title: Circulating Pneumolysin Is a Potent Inducer of Cardiac Injury during Pneumococcal Infection

    doi: 10.1371/journal.ppat.1004836

    Figure Lengend Snippet: PLY induces endoplasmic reticulum (ER) stress pathway without progressing to apoptosis in HL-1 cardiomyocytes. ( A ) Typical Western blots showing the activation of ER stress markers, (p-elF2α, p-IRE, BiP), JNK and ERK in HL-1 cells 30 min after PLY treatment. ( B ) Representative western blots showing the effect of PLY on the induction of apoptotic markers CHOP and active caspase-3 after 8 hour of treatment. Thapsigargin (Tg 5 μM) and Staurosporin (Stau 10 μM) were used as positive inducers of CHOP and active caspase-3 respectively. ( C-G ) Effects of PLY (1.0 μg/ml) and PLY+ 4-Phenylbutyric acid (4-PBA, an ER stress inhibitor, 10 mM) on Peak Shortening (C), (+dL/dt) (D), TTP (E), tR 90 (F) and (-dL/dt) (G) of HL-1 cells after 30 min treatment are presented as Mean±SD (n = 9 from 3 independent experiments). * p

    Article Snippet: Pathological examination of murine heart sections Hematoxylin and eosin (H & E) and immunohistochemical staining with anti-active caspase-3 (abcam) (1:10) or anti-serotype 1, 2, 4, 5 and 18 pneumococcus antiserum (SSI Diagnostica) (1:500) were carried out as described previously [ ].

    Techniques: Western Blot, Activation Assay

    The mRNA ( A ) and protein ( B ) expressions of caspase-3 caspase-8 and caspase-9 in human oral squamous carcinoma HSC-3 cells. * p

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    Article Title: Lactobacillus casei Strain Shirota Enhances the In Vitro Antiproliferative Effect of Geniposide in Human Oral Squamous Carcinoma HSC-3 Cells

    doi: 10.3390/molecules23051069

    Figure Lengend Snippet: The mRNA ( A ) and protein ( B ) expressions of caspase-3 caspase-8 and caspase-9 in human oral squamous carcinoma HSC-3 cells. * p

    Article Snippet: Every group was isolated for 2 h using non-fat milk sealing liquid (5%), and thereafter, it was combined with primary antibody of caspase-3 (No. PA5-16332, 1:500 dilution, Thermo Fisher Scientific, Waltham, MA, USA), caspase-8 (No. MA5-11558, 1:2000 dilution, Thermo Fisher Scientific), caspase-9 (No. PA5-16355, 1:500 dilution, Thermo Fisher Scientific), Bax (No. MA5-14003, 1:100 dilution, Thermo Fisher Scientific), Bcl-2 (No. MA5-11757, 1:400 dilution, Thermo Fisher Scientific), Bcl-xL (No. MA5-15142, 1:1000 dilution, Thermo Fisher Scientific), Fas (No. ab82419, 1:1000 dilution, Abcam, Cambridge, MA, USA), FasL (No. ab15285, 1:1000 dilution, Abcam), p53 (No. MA5-12557, 1:200 dilution, Thermo Fisher Scientific), p21 (No. MA5-14949, 1:1000 dilution, Thermo Fisher Scientific), HIAP-1 (No. PA1-26473, 1:200 dilution, Thermo Fisher Scientific), HIAP-2 (No. PA5-47036, 1:400 dilution, Thermo Fisher Scientific), NF-κB (No. ab195854, 1:2000 dilution, Abcam), IκB-α (No. ab7217, 1:2000 dilution, Abcam), COX-2 (No. 35-8200, 1:500 dilution, Thermo Fisher Scientific), iNOS (No. PA3-030A, 1:2000 dilution, Thermo Fisher Scientific), MMP-2 (No. 436000, 1:500 dilution, Thermo Fisher Scientific), MMP-9 (No. MA5-15886, 1:1000 dilution, Thermo Fisher Scientific), TIMP-1 (No. MA1-773, 1:500 dilution, Thermo Fisher Scientific), TIMP-2 (No. MA5-12207, 1:200 dilution, Thermo Fisher Scientific) and β-actin (No. MA1-140, 1:500 dilution) at 4 °C for 12 h. Individually, each group was mixed with secondary antibody after being washed with TBST three times and incubated by shaking at 25 °C for 2 h, followed by rinsing with TBST three times.

    Techniques:

    Substitution of Matrigel/rCOL by hCOL from fibroblasts leads to lumenized EC network formation with involvement of apoptosis. ( a , b ) 3D hydrogel constructs containing GFP positive HUVECs and hASCs and hCOL after 14 days of cultivation in SFM. ( c ) Virtual stacks of GFP-HUVECs co-cultured with hASCs in hCOL for 14 days and stained for α-SMA. Nuclei were counterstained with DAPI. α-SMA positive cells are in physical contact with GFP-HUVECs. ( d ) Virtual stacks of HUVECs co-cultured with hASCs and incubated with Texas red-labeled Dextran. ( e – e ”’) Images of cryo-sections from constructs containing GFP-HUVECs, hASCs and hCOL. Constructs were cultivated for 14 days in SFM and incubated with Texas red-labeled dextran. Cryo-sections were stained with DAPI. ( e ) GFP-HUVEC, ( e ’) DAPI, ( e ”) Texas red-labeled dextran, ( e ”’) Superimposed image of ( e , e ’, e ”). Intense DAPI staining of fragmented DNA is localized within the lumen filled with Texas red-labeled dextran. ( f – f ”’) Images of cryo-sections from constructs containing GFP-HUVECs, hASCs and hCOL. Constructs were cultivated for 14 days in SFM. Cryo-sections were stained for cleaved caspase 3 and DAPI. ( f ) GFP-HUVEC, ( f ’) DAPI, ( f ”) cleaved caspase 3 (CASP3), ( f ”’) superimposed image of ( f , f ’, f ”). Intense DAPI staining of fragmented DNA is localized within the lumen and co-localizes with the signal for cleaved caspase 3. Scale bar: ( a ) 500 µm; ( b ) 200 µm; ( c – f ”’) 30 µm.

    Journal: Scientific Reports

    Article Title: Formation of three-dimensional tubular endothelial cell networks under defined serum-free cell culture conditions in human collagen hydrogels

    doi: 10.1038/s41598-019-41985-6

    Figure Lengend Snippet: Substitution of Matrigel/rCOL by hCOL from fibroblasts leads to lumenized EC network formation with involvement of apoptosis. ( a , b ) 3D hydrogel constructs containing GFP positive HUVECs and hASCs and hCOL after 14 days of cultivation in SFM. ( c ) Virtual stacks of GFP-HUVECs co-cultured with hASCs in hCOL for 14 days and stained for α-SMA. Nuclei were counterstained with DAPI. α-SMA positive cells are in physical contact with GFP-HUVECs. ( d ) Virtual stacks of HUVECs co-cultured with hASCs and incubated with Texas red-labeled Dextran. ( e – e ”’) Images of cryo-sections from constructs containing GFP-HUVECs, hASCs and hCOL. Constructs were cultivated for 14 days in SFM and incubated with Texas red-labeled dextran. Cryo-sections were stained with DAPI. ( e ) GFP-HUVEC, ( e ’) DAPI, ( e ”) Texas red-labeled dextran, ( e ”’) Superimposed image of ( e , e ’, e ”). Intense DAPI staining of fragmented DNA is localized within the lumen filled with Texas red-labeled dextran. ( f – f ”’) Images of cryo-sections from constructs containing GFP-HUVECs, hASCs and hCOL. Constructs were cultivated for 14 days in SFM. Cryo-sections were stained for cleaved caspase 3 and DAPI. ( f ) GFP-HUVEC, ( f ’) DAPI, ( f ”) cleaved caspase 3 (CASP3), ( f ”’) superimposed image of ( f , f ’, f ”). Intense DAPI staining of fragmented DNA is localized within the lumen and co-localizes with the signal for cleaved caspase 3. Scale bar: ( a ) 500 µm; ( b ) 200 µm; ( c – f ”’) 30 µm.

    Article Snippet: Blocking was performed with 2% donkey serum in PBS for 20 minutes at room temperature followed by overnight incubation with primary antibody α-smooth muscle actin (α-SMA) (DAKO, 1:400) or cleaved caspase-3 (CASP3) (Cell Signaling Technology, 1:200) at 4 °C.

    Techniques: Construct, Cell Culture, Staining, Incubation, Labeling

    1, 25(OH)2 D3 induces apoptosis of PEL cells. PEL cells were cultured in the presence or absence of 1, 25(OH)2 D3 (10 nM). ( A ) After 48 h in culture, cells were washed and stained with Annexin V and PI and analyzed by flow cytometry. The numbers in the lower right quadrant represent the percent of apoptotic cells in culture. ( B and C ) . Cleaved caspase-3, and cleaved PARP expressions were detected by Western blot in JSC-1 and HBL-6 cells after treatment with 1, 25(OH)2 D3. β-actin was used to normalize protein loading.

    Journal: Scientific Reports

    Article Title: 1, 25(OH)2 D3 Induces Reactivation and Death of Kaposi’s Sarcoma-Associated Herpesvirus of Primary Effusion Lymphoma cells

    doi: 10.1038/s41598-017-12676-x

    Figure Lengend Snippet: 1, 25(OH)2 D3 induces apoptosis of PEL cells. PEL cells were cultured in the presence or absence of 1, 25(OH)2 D3 (10 nM). ( A ) After 48 h in culture, cells were washed and stained with Annexin V and PI and analyzed by flow cytometry. The numbers in the lower right quadrant represent the percent of apoptotic cells in culture. ( B and C ) . Cleaved caspase-3, and cleaved PARP expressions were detected by Western blot in JSC-1 and HBL-6 cells after treatment with 1, 25(OH)2 D3. β-actin was used to normalize protein loading.

    Article Snippet: The phospho-p38 mitogen-activated protein kinase (MAPK), LANA and caspase-3 antibodies were purchased from Imgenex.

    Techniques: Cell Culture, Staining, Flow Cytometry, Cytometry, Western Blot

    Effects of caspases inhibition by Z-VAD-FMK on apoptosis in PEL cells. JSC-1 cells were pre-treated with the caspase inhibitor Z-VAD-FMK (20 µM) for 2 h, followed by treatment with 10 nM 1, 25(OH)2 D3 for 24 h. ( A ) Cells were stained with Annexin V/PI, and apoptosis was determined using flow cytometry. Values for cells treated with DIM and Z-VAD-FMK were significantly reduced as compared to treatment with 1, 25(OH)2 D3 alone. Total protein extracts were prepared and subjected to Western blot assay for cleaved PARP and cleaved caspase-3 ( B ). Cell lysates collected 24 h after induction with the 1, 25(OH)2 D3 (10 nm) were immunoblotted with anti-ORF57 and anti K8.1 antibody. Effect of Caspase inhibitor on the expression of ORF57 or K8.1 in cells treated with 1, 25(OH)2 D3 alone or both 1, 25(OH)2 D3 and Z-VAD-FMK ( C and D ). β-actin was used to normaliz protein loading.

    Journal: Scientific Reports

    Article Title: 1, 25(OH)2 D3 Induces Reactivation and Death of Kaposi’s Sarcoma-Associated Herpesvirus of Primary Effusion Lymphoma cells

    doi: 10.1038/s41598-017-12676-x

    Figure Lengend Snippet: Effects of caspases inhibition by Z-VAD-FMK on apoptosis in PEL cells. JSC-1 cells were pre-treated with the caspase inhibitor Z-VAD-FMK (20 µM) for 2 h, followed by treatment with 10 nM 1, 25(OH)2 D3 for 24 h. ( A ) Cells were stained with Annexin V/PI, and apoptosis was determined using flow cytometry. Values for cells treated with DIM and Z-VAD-FMK were significantly reduced as compared to treatment with 1, 25(OH)2 D3 alone. Total protein extracts were prepared and subjected to Western blot assay for cleaved PARP and cleaved caspase-3 ( B ). Cell lysates collected 24 h after induction with the 1, 25(OH)2 D3 (10 nm) were immunoblotted with anti-ORF57 and anti K8.1 antibody. Effect of Caspase inhibitor on the expression of ORF57 or K8.1 in cells treated with 1, 25(OH)2 D3 alone or both 1, 25(OH)2 D3 and Z-VAD-FMK ( C and D ). β-actin was used to normaliz protein loading.

    Article Snippet: The phospho-p38 mitogen-activated protein kinase (MAPK), LANA and caspase-3 antibodies were purchased from Imgenex.

    Techniques: Inhibition, Staining, Flow Cytometry, Cytometry, Western Blot, Expressing

    VDR knockdown inhibits KSHV reactivation. JSC-1 cells were transduced with lentiviral particles containing either scrambled shRNA or shRNAs directed against the human VDR transcript. ( A ) Western blot analysis of stably transduced cells shows near complete absence of VDR protein in JSC-1 cells. ( B and C ) Cell apoptosis was significantly decreased in JSC-1 cells with VDR knockdown as compared to control shRNA transduced cells. ( D ) Western blotting with anti caspase-3, anti ORF57 and β-actin antibodies of cell lysates of JSC-1 VDR KO cell line and JSC-1 Ctrl treated with 10 nM of 1, 25(OH)2 D3 and 20 ng/ml of TPA used as positive control.

    Journal: Scientific Reports

    Article Title: 1, 25(OH)2 D3 Induces Reactivation and Death of Kaposi’s Sarcoma-Associated Herpesvirus of Primary Effusion Lymphoma cells

    doi: 10.1038/s41598-017-12676-x

    Figure Lengend Snippet: VDR knockdown inhibits KSHV reactivation. JSC-1 cells were transduced with lentiviral particles containing either scrambled shRNA or shRNAs directed against the human VDR transcript. ( A ) Western blot analysis of stably transduced cells shows near complete absence of VDR protein in JSC-1 cells. ( B and C ) Cell apoptosis was significantly decreased in JSC-1 cells with VDR knockdown as compared to control shRNA transduced cells. ( D ) Western blotting with anti caspase-3, anti ORF57 and β-actin antibodies of cell lysates of JSC-1 VDR KO cell line and JSC-1 Ctrl treated with 10 nM of 1, 25(OH)2 D3 and 20 ng/ml of TPA used as positive control.

    Article Snippet: The phospho-p38 mitogen-activated protein kinase (MAPK), LANA and caspase-3 antibodies were purchased from Imgenex.

    Techniques: Transduction, shRNA, Western Blot, Stable Transfection, Positive Control

    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:

    Western blot analysis of Cx43, HC1, and Casp3 expression from astrocytes following OGD treatment. Astrocytes were treated with OGD techniques. The lysates were resolved by SDS-PAGE, and immunopositive bands were semiquantified by scanning laser densitometry. Data are expressed as mean ± SD corresponding to data displayed, * P

    Journal: Child neurology open

    Article Title: Connexin 43 and Its Hemichannels Mediate Hypoxia–Ischemia-Induced Cell Death in Neonatal Rats

    doi: 10.1177/2329048X14544955

    Figure Lengend Snippet: Western blot analysis of Cx43, HC1, and Casp3 expression from astrocytes following OGD treatment. Astrocytes were treated with OGD techniques. The lysates were resolved by SDS-PAGE, and immunopositive bands were semiquantified by scanning laser densitometry. Data are expressed as mean ± SD corresponding to data displayed, * P

    Article Snippet: Rabbit caspase 3 (Casp3) antibody was from Sigma-Aldrich (HPA002643; Poole).

    Techniques: Western Blot, Expressing, SDS Page

    Western blot analysis of Cx43, HC1, and Casp3 expression in subventricular zone of rat’s brain after HI insult. A, Protein expression of Cx43, HC1, Casp3, and tublin was identified from the tissue extracts in subventricular zone of rat’s brain. B, Bar histograms show the mean ± SD corresponding to data displayed in part A. * P

    Journal: Child neurology open

    Article Title: Connexin 43 and Its Hemichannels Mediate Hypoxia–Ischemia-Induced Cell Death in Neonatal Rats

    doi: 10.1177/2329048X14544955

    Figure Lengend Snippet: Western blot analysis of Cx43, HC1, and Casp3 expression in subventricular zone of rat’s brain after HI insult. A, Protein expression of Cx43, HC1, Casp3, and tublin was identified from the tissue extracts in subventricular zone of rat’s brain. B, Bar histograms show the mean ± SD corresponding to data displayed in part A. * P

    Article Snippet: Rabbit caspase 3 (Casp3) antibody was from Sigma-Aldrich (HPA002643; Poole).

    Techniques: Western Blot, Expressing

    Immunostaining for the expression of Cx43, HC1, and Casp3 in rat’s brain after HI insult. Cx43, HC1, and Casp3 were stained green, and nuclei were stained blue. Bar = 25 µm. a1-3, The expression of Cx43, HC1, and Casp3 in SVZ from rat without the insult of HI (Cont.); b1-3, The expression of Cx43, HC1, and Casp3 in SVZ from rat 1 hour after HI (T1 hour); c1-3, The expression of Cx43, HC1, and Casp3 in SVZ from rat 12 hours after HI (T12 hour); d1-3, The expression of Cx43, HC1, and Casp3 from rat 24 hours after HI (T24 hour); e1-3, The expression of Cx43, HC1, and Casp3 in SVZ from rat 48 hours after HI (T48 hour); f1, The plaque area (pixels) of Cx43 and HC1 staining per cell with the time points. f2, The percentage of Casp3 positive cells (%) with the time points. * P

    Journal: Child neurology open

    Article Title: Connexin 43 and Its Hemichannels Mediate Hypoxia–Ischemia-Induced Cell Death in Neonatal Rats

    doi: 10.1177/2329048X14544955

    Figure Lengend Snippet: Immunostaining for the expression of Cx43, HC1, and Casp3 in rat’s brain after HI insult. Cx43, HC1, and Casp3 were stained green, and nuclei were stained blue. Bar = 25 µm. a1-3, The expression of Cx43, HC1, and Casp3 in SVZ from rat without the insult of HI (Cont.); b1-3, The expression of Cx43, HC1, and Casp3 in SVZ from rat 1 hour after HI (T1 hour); c1-3, The expression of Cx43, HC1, and Casp3 in SVZ from rat 12 hours after HI (T12 hour); d1-3, The expression of Cx43, HC1, and Casp3 from rat 24 hours after HI (T24 hour); e1-3, The expression of Cx43, HC1, and Casp3 in SVZ from rat 48 hours after HI (T48 hour); f1, The plaque area (pixels) of Cx43 and HC1 staining per cell with the time points. f2, The percentage of Casp3 positive cells (%) with the time points. * P

    Article Snippet: Rabbit caspase 3 (Casp3) antibody was from Sigma-Aldrich (HPA002643; Poole).

    Techniques: Immunostaining, Expressing, Staining

    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

    When neither CTSB deletion nor pH neutralization affected necrosis, we investigated caspase 3 activation and apoptosis at the time point of maximal protease activation in acini following CCK stimulation (30 min). A , pH neutralization with chloroquine

    Journal: The Journal of Biological Chemistry

    Article Title: Cathepsin B Activity Initiates Apoptosis via Digestive Protease Activation in Pancreatic Acinar Cells and Experimental Pancreatitis *

    doi: 10.1074/jbc.M116.718999

    Figure Lengend Snippet: When neither CTSB deletion nor pH neutralization affected necrosis, we investigated caspase 3 activation and apoptosis at the time point of maximal protease activation in acini following CCK stimulation (30 min). A , pH neutralization with chloroquine

    Article Snippet: Nafamostat, a trypsin inhibitor, recombinant human caspase 3, as well as chloroquine were from Sigma.

    Techniques: Neutralization, Activation Assay

    A–C , to study whether trypsin-induced caspase 3 activation translates into apoptosis in vivo , we used two models of experimental pancreatitis and measured apoptosis by TUNEL assay in tissue sections. After 8 h of caerulein-induced pancreatitis,

    Journal: The Journal of Biological Chemistry

    Article Title: Cathepsin B Activity Initiates Apoptosis via Digestive Protease Activation in Pancreatic Acinar Cells and Experimental Pancreatitis *

    doi: 10.1074/jbc.M116.718999

    Figure Lengend Snippet: A–C , to study whether trypsin-induced caspase 3 activation translates into apoptosis in vivo , we used two models of experimental pancreatitis and measured apoptosis by TUNEL assay in tissue sections. After 8 h of caerulein-induced pancreatitis,

    Article Snippet: Nafamostat, a trypsin inhibitor, recombinant human caspase 3, as well as chloroquine were from Sigma.

    Techniques: Activation Assay, In Vivo, TUNEL Assay

    Nickel exposure induced variation of caspase-3 (A) and caspase-9 (B) activities in piscine brain exposed to 0% (0.00 mg L -1 ), 10% (4.1 mg L -1 ) and 20% (8.2 mg L -1 ) of 96 h LC 50 of Ni at 20, 40 and 60 days of exposure. Data are mean ± SE of eight observations. Results of DMR Test have been represented by small letters. Common letter between any two bars indicates their similarity while two different letters indicate significant difference at 5% level.

    Journal: Interdisciplinary Toxicology

    Article Title: Mitochondrial respiratory chain inhibition and Na+K+ATPase dysfunction are determinant factors modulating the toxicity of nickel in the brain of indian catfish Clarias batrachus L.

    doi: 10.2478/intox-2018-0030

    Figure Lengend Snippet: Nickel exposure induced variation of caspase-3 (A) and caspase-9 (B) activities in piscine brain exposed to 0% (0.00 mg L -1 ), 10% (4.1 mg L -1 ) and 20% (8.2 mg L -1 ) of 96 h LC 50 of Ni at 20, 40 and 60 days of exposure. Data are mean ± SE of eight observations. Results of DMR Test have been represented by small letters. Common letter between any two bars indicates their similarity while two different letters indicate significant difference at 5% level.

    Article Snippet: Assessment of caspase-3 and caspase-9 activities Caspase-3 and caspase-9 assays were performed on the brain samples according to the manufacturer’s protocol (Biovision, K106-100 and K119-100).

    Techniques:

    A and B show percent staining for Ki67 and Mcm-2 with 95% confidence intervals across progressive tissue compartments. There is a strong shift in proliferation from basal to luminal cell compartments in both markers. C and D show a-casp3 and Bcl-2 scores with 95% confidence intervals. Only the luminal compartment was scored for these 2 markers. For a-casp3 a sharp significant drop in expression was seen in HGPIN and cancer compartments. No differential trend was observed for Bcl-2. Supernormal glands showed significantly higher luminal Mcm-2 indices and higher a-casp-3 activity than normal glands suggestive of a field effect.

    Journal: BMC Cancer

    Article Title: Alteration of proliferation and apoptotic markers in normal and premalignant tissue associated with prostate cancer

    doi: 10.1186/1471-2407-6-73

    Figure Lengend Snippet: A and B show percent staining for Ki67 and Mcm-2 with 95% confidence intervals across progressive tissue compartments. There is a strong shift in proliferation from basal to luminal cell compartments in both markers. C and D show a-casp3 and Bcl-2 scores with 95% confidence intervals. Only the luminal compartment was scored for these 2 markers. For a-casp3 a sharp significant drop in expression was seen in HGPIN and cancer compartments. No differential trend was observed for Bcl-2. Supernormal glands showed significantly higher luminal Mcm-2 indices and higher a-casp-3 activity than normal glands suggestive of a field effect.

    Article Snippet: For a-casp3 and Bcl-2, a mean of 15 (1–33) 40 × fields was scored.

    Techniques: Staining, Expressing, Activity Assay

    Figure 1 shows the expression of proliferation and apoptotic markers in the various compartments of the prostate. The immunomarkers are shown in columns from left to right (Ki67, Mcm-2, a-Casp3, and Bcl-2) and the compartments are shown in rows from top to bottom (Normal, HGPIN and IGCA). Normal compartment showed predominant basal cell (green arrows) positivity for Ki67 and Mcm-2. A shift towards the luminal compartment (red arrows) is noted for these two proliferation markers in HGPIN. Activated Caspase-3 on the other hand, showed a progressive decrease in the luminal cell staining from normal through PIN to cancer. Bcl-2 did not show any differential trend across compartments. Note that the strongly positive basal cells (for Bcl-2) serve as positive internal controls for the assay.

    Journal: BMC Cancer

    Article Title: Alteration of proliferation and apoptotic markers in normal and premalignant tissue associated with prostate cancer

    doi: 10.1186/1471-2407-6-73

    Figure Lengend Snippet: Figure 1 shows the expression of proliferation and apoptotic markers in the various compartments of the prostate. The immunomarkers are shown in columns from left to right (Ki67, Mcm-2, a-Casp3, and Bcl-2) and the compartments are shown in rows from top to bottom (Normal, HGPIN and IGCA). Normal compartment showed predominant basal cell (green arrows) positivity for Ki67 and Mcm-2. A shift towards the luminal compartment (red arrows) is noted for these two proliferation markers in HGPIN. Activated Caspase-3 on the other hand, showed a progressive decrease in the luminal cell staining from normal through PIN to cancer. Bcl-2 did not show any differential trend across compartments. Note that the strongly positive basal cells (for Bcl-2) serve as positive internal controls for the assay.

    Article Snippet: For a-casp3 and Bcl-2, a mean of 15 (1–33) 40 × fields was scored.

    Techniques: Expressing, Staining

    BSO decreased cell proliferation and induced cell apoptosis in ESCC. ( a ) PCNA IHC for cell proliferation in control and BSO-treated ESCC. ( b ) Quantification of cell proliferation in control and BSO-treated ESCC. ( c ) Cleaved caspase3 indicating cell apoptosis in control and BSO-treated ESCC. ( d ) Quantification of cell apoptosis in control and BSO-treated ESCC. IHC, immunohistochemical.

    Journal: Experimental & Molecular Medicine

    Article Title: Inhibition of glutathione metabolism attenuates esophageal cancer progression

    doi: 10.1038/emm.2017.15

    Figure Lengend Snippet: BSO decreased cell proliferation and induced cell apoptosis in ESCC. ( a ) PCNA IHC for cell proliferation in control and BSO-treated ESCC. ( b ) Quantification of cell proliferation in control and BSO-treated ESCC. ( c ) Cleaved caspase3 indicating cell apoptosis in control and BSO-treated ESCC. ( d ) Quantification of cell apoptosis in control and BSO-treated ESCC. IHC, immunohistochemical.

    Article Snippet: Immunohistochemistry Paraffin sections of ESCC tissue samples from mice were antigen retrieved, blocked and processed as previously described., Primary antibodies to mouse K14 (ab7800, 1:500), p63 (ab53039, 1:100), PCNA (ab29, 1:200) and cleaved Caspase3 (ab13847, 1:100) were purchased from Abcam.

    Techniques: Immunohistochemistry

    FKBP11 and GRP78 expression levels are increased in TNBS-induced colitis. Western blot analyses revealed that the expression levels of (A) FKBP11 and GRP78, and (B) active caspase-3 in TNBS induced colitis are significantly enhanced 3 days post-treatment compared with the ETOH group. Bar graphs demonstrate the quantitative analysis of FKBP11, GRP78 and active caspase-3 vs. GAPDH. (C) Immunohistochemistry analysis of FKBP11 and GRP78 expression in colonic mucosa of mice treated with TNBS for 3 days (scale bar, 50 µm). Data are presented as mean ± standard error (n=3). *P

    Journal: Molecular Medicine Reports

    Article Title: FKBP11 protects intestinal epithelial cells against inflammation-induced apoptosis via the JNK-caspase pathway in Crohn's disease

    doi: 10.3892/mmr.2018.9485

    Figure Lengend Snippet: FKBP11 and GRP78 expression levels are increased in TNBS-induced colitis. Western blot analyses revealed that the expression levels of (A) FKBP11 and GRP78, and (B) active caspase-3 in TNBS induced colitis are significantly enhanced 3 days post-treatment compared with the ETOH group. Bar graphs demonstrate the quantitative analysis of FKBP11, GRP78 and active caspase-3 vs. GAPDH. (C) Immunohistochemistry analysis of FKBP11 and GRP78 expression in colonic mucosa of mice treated with TNBS for 3 days (scale bar, 50 µm). Data are presented as mean ± standard error (n=3). *P

    Article Snippet: The membranes were blocked with 5% non-fat milk for 1 h at room temperature and incubated overnight at 4°C with the following primary antibodies: FK506 binding protein 11 (FKBP11, goat; 1:500; cat. no. AP6790a; Abgent, Inc., San Diego, CA, USA), 78 kDa glucose-regulated protein (GRP78, mouse; 1:500; cat. no. sc-13539), Bcl2 associated X apoptosis regulator (Bax, rabbit; 1:500; cat. no. sc-20067), proliferating cell nuclear antigen (PCNA, mouse; 1:500; cat. no. sc-25280; all Santa Cruz Biotechnology, Inc., Dallas, TX, USA), active caspase-3 (rabbit; 1:500; cat. no. AP3725a; Abgent, Inc.), c-Jun N-terminal kinase (JNK, mouse; 1:500; cat. no. sc-7345), phosphorylated JNK (p-JNK, mouse; 1:500), caspase-4 (human; 1:500; cat. no. sc-56056), caspase-12 (human; 1:500; sc56056), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, rabbit; 1:1,000; cat. no. sc-32233; all Santa Cruz Biotechnology, Inc.).

    Techniques: Expressing, Western Blot, Immunohistochemistry, Mouse Assay

    FKBP11 is involved in IFN-γ/TNF-α-induced apoptosis in HT-29 cells. (A) HT-29 cells were transfected with either the FKBP11 overexpressing plasmid pCMV-HA-FKBP11, blank control plasmid pCMV-HA, controls siRNA or FKBP11siRNA for 48 h and then treated with IFN-γ (2.5 ng/ml) and TNF-α (50 ng/ml) for 24 h. Western blot analyses detected FKBP11, BAX and active caspase-3 expression levels following treatment with IFN-γ/TNF-α compared with the untreated control and control plasmid (HA). # P

    Journal: Molecular Medicine Reports

    Article Title: FKBP11 protects intestinal epithelial cells against inflammation-induced apoptosis via the JNK-caspase pathway in Crohn's disease

    doi: 10.3892/mmr.2018.9485

    Figure Lengend Snippet: FKBP11 is involved in IFN-γ/TNF-α-induced apoptosis in HT-29 cells. (A) HT-29 cells were transfected with either the FKBP11 overexpressing plasmid pCMV-HA-FKBP11, blank control plasmid pCMV-HA, controls siRNA or FKBP11siRNA for 48 h and then treated with IFN-γ (2.5 ng/ml) and TNF-α (50 ng/ml) for 24 h. Western blot analyses detected FKBP11, BAX and active caspase-3 expression levels following treatment with IFN-γ/TNF-α compared with the untreated control and control plasmid (HA). # P

    Article Snippet: The membranes were blocked with 5% non-fat milk for 1 h at room temperature and incubated overnight at 4°C with the following primary antibodies: FK506 binding protein 11 (FKBP11, goat; 1:500; cat. no. AP6790a; Abgent, Inc., San Diego, CA, USA), 78 kDa glucose-regulated protein (GRP78, mouse; 1:500; cat. no. sc-13539), Bcl2 associated X apoptosis regulator (Bax, rabbit; 1:500; cat. no. sc-20067), proliferating cell nuclear antigen (PCNA, mouse; 1:500; cat. no. sc-25280; all Santa Cruz Biotechnology, Inc., Dallas, TX, USA), active caspase-3 (rabbit; 1:500; cat. no. AP3725a; Abgent, Inc.), c-Jun N-terminal kinase (JNK, mouse; 1:500; cat. no. sc-7345), phosphorylated JNK (p-JNK, mouse; 1:500), caspase-4 (human; 1:500; cat. no. sc-56056), caspase-12 (human; 1:500; sc56056), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, rabbit; 1:1,000; cat. no. sc-32233; all Santa Cruz Biotechnology, Inc.).

    Techniques: Transfection, Plasmid Preparation, Western Blot, Expressing

    Expression of FKBP11 and GRP78 are increased in intestinal tissues of patients with CD. Western blot analyses of (A) FKBP11 and GRP78 expressions levels, and (B) active caspase-3 expression levels in the intestinal tissues of patients with active CD and control tissues. GAPDH was used as a loading control. Bar graphs demonstrate the semi-quantitative analysis of FKBP11, GRP78 and active caspase-3 protein expression levels vs. GAPDH. (C) Immunohistochemistry analysis of FKBP11 and GRP78 expressions in mucosal biopsies tissues obtained from inflamed tissues with active CD and control tissues. Scale bar, 50 µm. Data are presented as mean ± standard error (n=3). *P

    Journal: Molecular Medicine Reports

    Article Title: FKBP11 protects intestinal epithelial cells against inflammation-induced apoptosis via the JNK-caspase pathway in Crohn's disease

    doi: 10.3892/mmr.2018.9485

    Figure Lengend Snippet: Expression of FKBP11 and GRP78 are increased in intestinal tissues of patients with CD. Western blot analyses of (A) FKBP11 and GRP78 expressions levels, and (B) active caspase-3 expression levels in the intestinal tissues of patients with active CD and control tissues. GAPDH was used as a loading control. Bar graphs demonstrate the semi-quantitative analysis of FKBP11, GRP78 and active caspase-3 protein expression levels vs. GAPDH. (C) Immunohistochemistry analysis of FKBP11 and GRP78 expressions in mucosal biopsies tissues obtained from inflamed tissues with active CD and control tissues. Scale bar, 50 µm. Data are presented as mean ± standard error (n=3). *P

    Article Snippet: The membranes were blocked with 5% non-fat milk for 1 h at room temperature and incubated overnight at 4°C with the following primary antibodies: FK506 binding protein 11 (FKBP11, goat; 1:500; cat. no. AP6790a; Abgent, Inc., San Diego, CA, USA), 78 kDa glucose-regulated protein (GRP78, mouse; 1:500; cat. no. sc-13539), Bcl2 associated X apoptosis regulator (Bax, rabbit; 1:500; cat. no. sc-20067), proliferating cell nuclear antigen (PCNA, mouse; 1:500; cat. no. sc-25280; all Santa Cruz Biotechnology, Inc., Dallas, TX, USA), active caspase-3 (rabbit; 1:500; cat. no. AP3725a; Abgent, Inc.), c-Jun N-terminal kinase (JNK, mouse; 1:500; cat. no. sc-7345), phosphorylated JNK (p-JNK, mouse; 1:500), caspase-4 (human; 1:500; cat. no. sc-56056), caspase-12 (human; 1:500; sc56056), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, rabbit; 1:1,000; cat. no. sc-32233; all Santa Cruz Biotechnology, Inc.).

    Techniques: Expressing, Western Blot, Immunohistochemistry

    HOXC 13 induces apoptosis of esophageal squamous cell carcinoma ( ESCC ) cells through regulating CASP 3. A, Gene ontology pathway analysis showed that genes co‐expressed with HOXC 13 were enriched in the “transcription,” “apoptotic process” and “proliferation” pathway. B,C, Quantitative RT ‐ PCR and western blot indicated that expression of CASP 3 was significantly upregulated by knockdown of HOXC 13. D,E, Z‐ DEVD ‐ FMK , a specific caspase‐3 inhibitor, partially reversed the inhibitory effect of sh‐ HOXC 13 on the proliferation of ECA 109 cells. F, Z‐ DEVD ‐ FMK partially reversed sh RNA ‐ HOXC 13‐induced apoptosis. G, Western blot showed that Z‐ DEVD ‐ FMK decreased the expression of CASP 3 and upregulated PARP , the enzyme digestion substrate of caspase‐3

    Journal: Cancer Science

    Article Title: HOXC13 promotes proliferation of esophageal squamous cell carcinoma via repressing transcription of CASP3, et al. HOXC13 promotes proliferation of esophageal squamous cell carcinoma via repressing transcription of CASP3

    doi: 10.1111/cas.13453

    Figure Lengend Snippet: HOXC 13 induces apoptosis of esophageal squamous cell carcinoma ( ESCC ) cells through regulating CASP 3. A, Gene ontology pathway analysis showed that genes co‐expressed with HOXC 13 were enriched in the “transcription,” “apoptotic process” and “proliferation” pathway. B,C, Quantitative RT ‐ PCR and western blot indicated that expression of CASP 3 was significantly upregulated by knockdown of HOXC 13. D,E, Z‐ DEVD ‐ FMK , a specific caspase‐3 inhibitor, partially reversed the inhibitory effect of sh‐ HOXC 13 on the proliferation of ECA 109 cells. F, Z‐ DEVD ‐ FMK partially reversed sh RNA ‐ HOXC 13‐induced apoptosis. G, Western blot showed that Z‐ DEVD ‐ FMK decreased the expression of CASP 3 and upregulated PARP , the enzyme digestion substrate of caspase‐3

    Article Snippet: Western blot showed that Z‐DEVD‐FMK decreased the expression of CASP3 and upregulated PARP, which is the digestion substrate of caspase‐3 (Figure G).

    Techniques: Quantitative RT-PCR, Western Blot, Expressing

    MiR‐503 inhibits tumor growth in vivo. A,B, Xenograft model in nude mice was used and tumor nodules harvested from miR‐503 group were smaller than miR‐nc group (C and D). Compared with the miR‐nc group, the miR‐503 group has reduced tumor volume and weight. E, Immunohistochemistry (IHC) analysis of xenograft tumors showed that HOXC13 protein level was decreased while CASP3 protein level was upregulated in the miR‐503 group

    Journal: Cancer Science

    Article Title: HOXC13 promotes proliferation of esophageal squamous cell carcinoma via repressing transcription of CASP3, et al. HOXC13 promotes proliferation of esophageal squamous cell carcinoma via repressing transcription of CASP3

    doi: 10.1111/cas.13453

    Figure Lengend Snippet: MiR‐503 inhibits tumor growth in vivo. A,B, Xenograft model in nude mice was used and tumor nodules harvested from miR‐503 group were smaller than miR‐nc group (C and D). Compared with the miR‐nc group, the miR‐503 group has reduced tumor volume and weight. E, Immunohistochemistry (IHC) analysis of xenograft tumors showed that HOXC13 protein level was decreased while CASP3 protein level was upregulated in the miR‐503 group

    Article Snippet: Western blot showed that Z‐DEVD‐FMK decreased the expression of CASP3 and upregulated PARP, which is the digestion substrate of caspase‐3 (Figure G).

    Techniques: In Vivo, Mouse Assay, Immunohistochemistry

    Autophagy inhibition leads to dysfunctional mitochondria and apoptosis.  a  Representative western blot of mitophagy markers from D2.0 R cells plated on BME or BME plus COL1 with or without HCQ on days 1 and 5.  b  Representative images of live D2.0 R cells transfected with the GFP-LC3 reporter and stained with MitoTracker® Red CMXRos on BME or BME plus COL1 matrices for 5 days. Scale bar is 20 µm  c  Mitochondrial (MitoTracker® Green), mitochondrial reactive oxygen species (ROS; MitoSox™) and mitochondrial membrane potential (TMRM) quantification in D2.0 R cells on BME or BME plus COL1 matrices with or without HCQ (mean ± s.e.m,  n  = 60,000 cells from 3 independent experiments. Comparisons by Mann–Whitney  U -test, two-sided. ** P  ≤ 0.01; **** P  ≤ 0.0001). NT, non-treated; HCQ-D5, hydroxychloroquine treatment beginning on day 5; MFI, mean fluorescence intensity.  d  MitoTempo is protective of ROS-induced cell death in D2.0 R cells on BME treated with HCQ. Cells were pre-treated for 5 days with 20 μM MitoTempo and subsequently exposed to 50 μM HCQ for 24 hrs. Cell viability was assessed by Cytotox Glo assay (left graph. Mean ± s.e.m,  n  = 3 wells. Comparisons by unpaired two-sided  T -test) and Caspase 3 and 7 activity (right graph. Mean ± s.e.m,  n  = 3 wells. Comparisons by unpaired two-sided  T -test). Data are representative of three independent experiments

    Journal: Nature Communications

    Article Title: Autophagy promotes the survival of dormant breast cancer cells and metastatic tumour recurrence

    doi: 10.1038/s41467-018-04070-6

    Figure Lengend Snippet: Autophagy inhibition leads to dysfunctional mitochondria and apoptosis. a Representative western blot of mitophagy markers from D2.0 R cells plated on BME or BME plus COL1 with or without HCQ on days 1 and 5. b Representative images of live D2.0 R cells transfected with the GFP-LC3 reporter and stained with MitoTracker® Red CMXRos on BME or BME plus COL1 matrices for 5 days. Scale bar is 20 µm c Mitochondrial (MitoTracker® Green), mitochondrial reactive oxygen species (ROS; MitoSox™) and mitochondrial membrane potential (TMRM) quantification in D2.0 R cells on BME or BME plus COL1 matrices with or without HCQ (mean ± s.e.m, n  = 60,000 cells from 3 independent experiments. Comparisons by Mann–Whitney U -test, two-sided. ** P  ≤ 0.01; **** P  ≤ 0.0001). NT, non-treated; HCQ-D5, hydroxychloroquine treatment beginning on day 5; MFI, mean fluorescence intensity. d MitoTempo is protective of ROS-induced cell death in D2.0 R cells on BME treated with HCQ. Cells were pre-treated for 5 days with 20 μM MitoTempo and subsequently exposed to 50 μM HCQ for 24 hrs. Cell viability was assessed by Cytotox Glo assay (left graph. Mean ± s.e.m, n  = 3 wells. Comparisons by unpaired two-sided T -test) and Caspase 3 and 7 activity (right graph. Mean ± s.e.m, n  = 3 wells. Comparisons by unpaired two-sided T -test). Data are representative of three independent experiments

    Article Snippet: Primary antibodies used were: from Cell Signaling Technology anti-ATG7 (8558, dilution 1/1000), p62 (5114, dilution 1/1000), BECN1 (3495, dilution 1/1000), LC3 (12741, dilution 1/1000), CASP3 (9662, dilution 1/1000), cleaved CASP3 (9661, dilution 1/1000), ɣH2Ax (p139S) (2577, dilution 1/1000), from Abcam anti-PINK1 (ab75487, dilution 1/500), MTOC-1 (ab14705, dilution 1/2000), TOM20 (ab56783, dilution 1/500) and from Thermo Scientific anti-Cyclophillin B (PA1-027A, dilution 1/300), followed by secondary antibodies (GE Healthcare).

    Techniques: Inhibition, Western Blot, Transfection, Staining, MANN-WHITNEY, Fluorescence, Glo Assay, Activity Assay