anti-cleaved caspase-3 Search Results


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  • 99
    Millipore cleaved caspase 3
    FOXD3 overexpression suppressed proliferation and enhanced starvation-induced apoptosis in U87MG cells. A: Western blotting analysis of FOXD3 and cleaved <t>caspase-3</t> expression in U87MG cells. B: Cell proliferation assay showing significantly enhanced proliferation rate of FOXD3-overexpressed U87MG cells in comparison with EV-treated U87MG cells ( P
    Cleaved Caspase 3, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 968 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc cleaved caspase 3
    Effect of VC on apoptosis. (A) Representative western blots and densitometric analysis for whole liver <t>caspase‐3</t> protein are shown. (B) TUNEL staining was performed as described in Materials and Methods for the 12‐week samples. Representative photomicrographs are shown (magnification ×200). (C) TUNEL‐positive hepatocytes and NPCs were quantified as described in Materials and Methods and are expressed as TUNEL‐positive cells per 1,000 hepatocytes. a P
    Cleaved Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 38294 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Cell Signaling Technology Inc anti cleaved caspase 3
    <t>Caspase</t> 3 staining in kidney sections and isolated podocytes. A) A Immunohistochemical staining with anti-caspase 3 antibody. Scale bar: 50μm (upper row); 20μm (lower row). B) Confocal micrographs from primary podocytes derived from described genotypes showing cytoplasm staining (GAPDH in green), apoptotic cells (Cleaved CASPASE-3 in red) and nuclei (DAPI in blue). Maximum intensity projection of Z-stacks is shown. Scale bars: 50 μm. OnOn the right panel, qquantification of caspase 3-positive cells. Data are presented as mean ± SD. Symbols * and § represent the significance levels vs. WT and Mpv17 -/- , respectively, calculated by one‐way ANOVA with Tukey’s post hoc multiple comparison test − : **** p
    Anti Cleaved Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 11253 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Cell Signaling Technology Inc rabbit anti casp3
    Premature apoptosis occured in C9ORF72 spinal MNs over ageing (a) Apoptosis in MAP2-positive neurons (in red) was revealed as cytosolic rim staining around Hoechst-positive nuclei (in blue) for cleaved caspase 3 <t>(Casp3,</t> in green) by confocal IF microscopy at D80 endpoints (fig. 3). Dotted boxed areas in image galleries are shown magnified on the right. Note the striking accumulation of Casp3 in parental C9 and C9-KO that was phenocopied by WT-KO. Conversely, apoptosis hardly occurred in C9-GC and parental Ctrl1. Scale bars = 10µm. (b) Quantification of (a), percentage of apoptotic cells in MAP2-positive population. Asterisks: highly significant increase in any pairwise comparison with unlabeled conditions, one-way ANOVA with Bonferroni post test, *P≤0.05, **P≤0.01, ***P≤0.001, N=60 images from 3 independent experiments, error bars=SD. All unlabeled conditions (i.e. with no asterisk) were not significantly different amongst themselves in any pairwise comparison. Remaining comparisons were: WT-KO vs C9: ***, WT-KO vs C9-KO: ***, C9 vs C9-KO: ***.
    Rabbit Anti Casp3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 88/100, based on 66 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc rabbit anti cleaved caspase 3
    Adeno-associated virus (AAV)-AEG-1 transduction of dopaminergic (DA) neurons in the in vivo SN of healthy mice. a Experimental schematic and the immunostaining for green fluorescent protein (GFP; green) and hemagglutinin (HA; brown) in the SNpc, which is outlined by the dotted elliptical shape, which was conducted following each viral injection. Scale bar, 200 μm. b Representative double immunofluorescent labeling of TH (red) and GFP (green) or TH and HA (green) in the SNpc. Scale bar, 20 μm. c Representative double immunofluorescent labeling for glial fibrillary acidic protein (GFAP)/ionized calcium binding adaptor molecule 1 (Iba1; red), which are markers of astrocytes and microglia, respectively, and GFP/HA (green) in the SNpc of healthy mice. Scale bar, 20 μm. d Immunostaining for TH in the SN and striatum (STR). Scale bars, 200 μm (black) and 50 μm (white) for the SN, and 1000 μm for the STR. e , f The number and optical density of the nigral TH-positive neurons and striatal TH-positive fibers, respectively (one-way ANOVA with Tukey’s post hoc test; n = 4 for each group). CON contralateral side, IPSI ipsilateral side. g Western blot analyses of the levels of cleaved <t>caspase-3</t> (c-caspase-3) and cleaved poly (ADP-ribose) polymerase 1 (c-PARP-1) following AEG-1 transduction in the SN of healthy brains. * p = 0.005 vs . CON (one-way ANOVA with Tukey’s post hoc test; n = 4 for each group)
    Rabbit Anti Cleaved Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 5764 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc rabbit anti cleaved caspase 3 antibody
    Apoptosis of MIN6 cells induced by TM is inhibited by overexpression of DHCR24. (a) A representative fluorescent image of cleaved <t>caspase-3</t> (green) was obtained by IC analysis 24 h after TM exposure. Scale bar, 50 μ m. (b) The relative fluorescence intensity of cleaved caspase-3 was analyzed with ImageJ software. Mean ± S.D. ( n = 3); ∗ p
    Rabbit Anti Cleaved Caspase 3 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 848 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti cleaved caspase 3
    miR-21 can reduce PC12 cell death and TRPM7 is a functional target of miR-21 (A) . MiR-21 transfection efficiency detected by qPCR (B) . Apoptosis rate of PC12 cells transfected with miR-21 mimics, miR-21 inhibitors, and their corresponding negative controls (NC) (C) . Fluo-4 assay of PC12 cells (D) . The expression level of TRPM7 in PC12 cells transfected with miR-21 mimics, miR-21 inhibitors, and their corresponding negative controls (NC) (E) . Target region of miR-21 and TRPM7 (F) . Luciferase assay for the combination of miR-21 and TRPM7 (G) . Western blot analyzing the expression level of TRPM7, p65, and p-IκB proteins (H) . Quantitative histogram of TRPM7 expression level in different groups (I) . Western blot analyzing the expression level of cleaved <t>caspase-3.</t> * p
    Anti Cleaved Caspase 3, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 818 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc rabbit polyclonal anti cleaved caspase 3
    Effects of CLU silencing on viability, proliferation/death and differentiation of cultured OA chondrocytes. ( A ) CLU mRNA and protein expression after 48 and 72 hours, respectively, in scramble (control) and siCLU transfected OA-HACs. ( B ) Cell viability by MTT assay after 24, 48 and 72 hours of scramble and siCLU transfected OA-HACs. ( C ) Representative immunocytochemical images and bar graphs showing Ki67 + and p21 + nuclei in cultured OA-HACs and ( D ) cleaved <t>caspase-3</t> + cytospinned OA-HACs after 72 hours of silencing. ( E ) Gene array shows the modulation of NFkB and Stat-3-related genes in siCLU transfected OA-HACs compared with scramble after 48 hours of silencing. Transcript levels were normalized on GAPDH (internal reference gene). ( F ) mRNA levels by Real-Time PCR of COL2A1, ACAN, MMP13, COL10A1, and ( G ) representative blot and densitometric analysis of MMP13 and COL10A1 in scramble and CLU-silenced OA-HACs. Data are expressed as mean values ± SEM of three independent experiments (each using pooled samples from n=3 OA donors) performed in triplicate. Abbreviations: OA-HACs, OA-Human articular chondrocytes; OD, optical density unit; RLU, relative light unit. Student’s t-test: * P
    Rabbit Polyclonal Anti Cleaved Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 674 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology anti cleaved caspase 3
    Percentage of live (Annexin V negative/PI negative cells), apoptotic (AnnexinV positive/PI negative cells) and dead (AnnexinV positive/PI positive cells or AnnexinV negative/PI positive cells) cells in LnCaP (A) and CW22rv1 (B) cell populations treated with continuous or pulsed Testosterone for 48 hours. The bars show the mean and SD of 36 replicates. Student-Newman-Keuls test for all pairwise comparisons in CW22rv1 and LnCaP cells at 48 hours after treatments. Expression levels, by western blot analysis, of <t>pro-caspase-3</t> and cleaved caspase3 in LnCaP (C) and CW22rv1 (D) tumor cells tumor cells treated with continuous or pulsed Testosterone for 48 hours.
    Anti Cleaved Caspase 3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 487 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc antibody against cleaved caspase 3
    Screening OB2C small molecule library for death ligands Cleaved <t>caspase</t> 3 positive beads (red arrow) in OB2C beads library. Positive beads were picked up for chemical decoding with automatic Edman microsequencing. ( a ) LLS1, ( b ) LLS2. Resynthesize death ligand LLS2 on beads for validation of death effect. ( c ) Cleaved caspase 3 staining.(100X) ( d ) Propidium Iodide staining for dead cells. (40X). Galectin-1 is a target protein of LLS2. (e) Eluted protein from pull-down assay was identified as galectin-1 by LC MS/MS. (f) Immunoblot analysis with anti-galectin-1 Ab, to validate the LC MS/MS result. 1: blank bead, 2: LLS2-beasd, 3: a irrelevant small molecule-bead control. (g) Immunostaining result reveals that LLS2 co-localizes with galectin-1. SKOV3, A549, PC3 and XPA3 cells were stained by biotinylated LLS2 (green), galectin-1 antibody (red), and nuclei were stained with DAPI (blue). (h) Computer modeling shows that LLS2 binds the interface of the galectin-1 dimer. Residues that are within 3.5 angstroms from LLS2 were shown. Scale bars: 50 μm.
    Antibody Against Cleaved Caspase 3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    FOXD3 overexpression suppressed proliferation and enhanced starvation-induced apoptosis in U87MG cells. A: Western blotting analysis of FOXD3 and cleaved caspase-3 expression in U87MG cells. B: Cell proliferation assay showing significantly enhanced proliferation rate of FOXD3-overexpressed U87MG cells in comparison with EV-treated U87MG cells ( P

    Journal: PLoS ONE

    Article Title: Decreased FOXD3 Expression Is Associated with Poor Prognosis in Patients with High-Grade Gliomas

    doi: 10.1371/journal.pone.0127976

    Figure Lengend Snippet: FOXD3 overexpression suppressed proliferation and enhanced starvation-induced apoptosis in U87MG cells. A: Western blotting analysis of FOXD3 and cleaved caspase-3 expression in U87MG cells. B: Cell proliferation assay showing significantly enhanced proliferation rate of FOXD3-overexpressed U87MG cells in comparison with EV-treated U87MG cells ( P

    Article Snippet: After blocking non-specific binding sites for 60 min with 8% non-fat milk, membranes were incubated overnight at 4°C with primary antibodies against Cleaved Caspase-3 (1:1,000; Millipore, Billerica, MA, USA), FOXD3 (1:1,000; Abcam, USA) or GAPDH (1:1,000; Cell Signaling Technology, USA).

    Techniques: Over Expression, Western Blot, Expressing, Proliferation Assay

    FOXD3 silencing promoted proliferation and inhibited starvation-induced apoptosis in SW1080 cells. A: Western blotting analysis of FOXD3 and cleaved caspase-3 expression in SW1080 cells. B: Cell proliferation assay showing significantly enhanced proliferation rate of FOXD3-silenced SW1080 cells in comparison with siNC-treated SW1080 cells ( P

    Journal: PLoS ONE

    Article Title: Decreased FOXD3 Expression Is Associated with Poor Prognosis in Patients with High-Grade Gliomas

    doi: 10.1371/journal.pone.0127976

    Figure Lengend Snippet: FOXD3 silencing promoted proliferation and inhibited starvation-induced apoptosis in SW1080 cells. A: Western blotting analysis of FOXD3 and cleaved caspase-3 expression in SW1080 cells. B: Cell proliferation assay showing significantly enhanced proliferation rate of FOXD3-silenced SW1080 cells in comparison with siNC-treated SW1080 cells ( P

    Article Snippet: After blocking non-specific binding sites for 60 min with 8% non-fat milk, membranes were incubated overnight at 4°C with primary antibodies against Cleaved Caspase-3 (1:1,000; Millipore, Billerica, MA, USA), FOXD3 (1:1,000; Abcam, USA) or GAPDH (1:1,000; Cell Signaling Technology, USA).

    Techniques: Western Blot, Expressing, Proliferation Assay

    Effect of VC on apoptosis. (A) Representative western blots and densitometric analysis for whole liver caspase‐3 protein are shown. (B) TUNEL staining was performed as described in Materials and Methods for the 12‐week samples. Representative photomicrographs are shown (magnification ×200). (C) TUNEL‐positive hepatocytes and NPCs were quantified as described in Materials and Methods and are expressed as TUNEL‐positive cells per 1,000 hepatocytes. a P

    Journal: Hepatology Communications

    Article Title: Vinyl chloride dysregulates metabolic homeostasis and enhances diet‐induced liver injury in mice

    doi: 10.1002/hep4.1151

    Figure Lengend Snippet: Effect of VC on apoptosis. (A) Representative western blots and densitometric analysis for whole liver caspase‐3 protein are shown. (B) TUNEL staining was performed as described in Materials and Methods for the 12‐week samples. Representative photomicrographs are shown (magnification ×200). (C) TUNEL‐positive hepatocytes and NPCs were quantified as described in Materials and Methods and are expressed as TUNEL‐positive cells per 1,000 hepatocytes. a P

    Article Snippet: Primary antibodies against caspase‐3, glyceraldehyde 3‐phospohate dehydrogenase (GAPDH) (Santa Cruz Biotechnology, Dallas, TX), cleaved caspase‐3 (Cell Signaling, Beverly, MA), and CCAT‐enhancer‐binding protein homologous protein (Chop), (Thermo Fisher Scientific) were used.

    Techniques: Western Blot, TUNEL Assay, Staining

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

    Journal: Cell Death & Disease

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

    doi: 10.1038/cddis.2014.514

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

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

    Techniques: Activity Assay, In Situ, Proximity Ligation Assay

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

    Journal: Cell Death & Disease

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

    doi: 10.1038/cddis.2014.514

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

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

    Techniques: Activation Assay

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

    Journal: Cell Death & Disease

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

    doi: 10.1038/cddis.2014.514

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

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

    Techniques: Activation Assay, Fractionation

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

    Journal: Cell Death & Disease

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

    doi: 10.1038/cddis.2014.514

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

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

    Techniques: Activation Assay, Expressing, FACS, Activity Assay

    Caspase 3 staining in kidney sections and isolated podocytes. A) A Immunohistochemical staining with anti-caspase 3 antibody. Scale bar: 50μm (upper row); 20μm (lower row). B) Confocal micrographs from primary podocytes derived from described genotypes showing cytoplasm staining (GAPDH in green), apoptotic cells (Cleaved CASPASE-3 in red) and nuclei (DAPI in blue). Maximum intensity projection of Z-stacks is shown. Scale bars: 50 μm. OnOn the right panel, qquantification of caspase 3-positive cells. Data are presented as mean ± SD. Symbols * and § represent the significance levels vs. WT and Mpv17 -/- , respectively, calculated by one‐way ANOVA with Tukey’s post hoc multiple comparison test − : **** p

    Journal: bioRxiv

    Article Title: Opa1 overexpression protects from early onset Mpv17-/--related mouse kidney disease

    doi: 10.1101/2020.03.18.996561

    Figure Lengend Snippet: Caspase 3 staining in kidney sections and isolated podocytes. A) A Immunohistochemical staining with anti-caspase 3 antibody. Scale bar: 50μm (upper row); 20μm (lower row). B) Confocal micrographs from primary podocytes derived from described genotypes showing cytoplasm staining (GAPDH in green), apoptotic cells (Cleaved CASPASE-3 in red) and nuclei (DAPI in blue). Maximum intensity projection of Z-stacks is shown. Scale bars: 50 μm. OnOn the right panel, qquantification of caspase 3-positive cells. Data are presented as mean ± SD. Symbols * and § represent the significance levels vs. WT and Mpv17 -/- , respectively, calculated by one‐way ANOVA with Tukey’s post hoc multiple comparison test − : **** p

    Article Snippet: Antibodies Mouse monoclonal anti-GAPDH (1:1,000; #ab8245; Abcam), rabbit monoclonal anti-TOM20 Alexa Fluor 647 (1:500; #ab209606; Abcam), Alexa Fluor 488 Goat anti-mouse (1:300; #A11004; Invitrogen), Alexa Fluor 647 Goat anti-rabbit (1:300; #A27040; Invitrogen), rabbit polyclonal anti-NPHS2 (1:300; #ab50339; Abcam), mouse monoclonal anti-CYTOKERATIN (1:300; #ab86734; Abcam), anti-cleaved caspase 3 (1:200; Cell Signalling, #9661).

    Techniques: Staining, Isolation, Immunohistochemistry, Derivative Assay

    dsRB-SCP/polyIC selectively induces apoptosis of PSMA-overexpressing cells A. Cells were seeded in triplicate, grown overnight, and treated with dsRB-SCP/polyIC, polyIC alone or dsRB-SCP alone, as indicated, for 100 h. Viability was quantified using the CellTiter-Glo Luminescent Cell Viability Assay (Promega). Results (mean and standard deviation) are representative of two independent experiments (**** P ≤ 0.0001, treatment with dsRB-SCP/polyIC of LNCaP vs PC3; **** P ≤ 0.0001 treatment of LNCaP with dsRB-SCP/polyIC vs polyIC alone). B. Surviving cells remained permanently arrested. Cells were seeded in triplicate, grown overnight, and treated as indicated. Medium was replaced and viability was quantified after 100/172/344 h using CellTiter-Glo (**** P ≤ 0.0001 dsRB-SCP/polyIC treatment vs UT). Control cells were unable to proliferate beyond 2.5 doublings because they had reached full confluence. C. LNCaP cells were treated for the indicated times with dsRB-SCP/ polyIC or polyIC alone, lysed and subjected to western blot analysis to detect full-length and cleaved Caspase-3 and PARP.

    Journal: Oncotarget

    Article Title: PSMA-homing dsRNA chimeric protein vector kills prostate cancer cells and activates anti-tumor bystander responses

    doi: 10.18632/oncotarget.15733

    Figure Lengend Snippet: dsRB-SCP/polyIC selectively induces apoptosis of PSMA-overexpressing cells A. Cells were seeded in triplicate, grown overnight, and treated with dsRB-SCP/polyIC, polyIC alone or dsRB-SCP alone, as indicated, for 100 h. Viability was quantified using the CellTiter-Glo Luminescent Cell Viability Assay (Promega). Results (mean and standard deviation) are representative of two independent experiments (**** P ≤ 0.0001, treatment with dsRB-SCP/polyIC of LNCaP vs PC3; **** P ≤ 0.0001 treatment of LNCaP with dsRB-SCP/polyIC vs polyIC alone). B. Surviving cells remained permanently arrested. Cells were seeded in triplicate, grown overnight, and treated as indicated. Medium was replaced and viability was quantified after 100/172/344 h using CellTiter-Glo (**** P ≤ 0.0001 dsRB-SCP/polyIC treatment vs UT). Control cells were unable to proliferate beyond 2.5 doublings because they had reached full confluence. C. LNCaP cells were treated for the indicated times with dsRB-SCP/ polyIC or polyIC alone, lysed and subjected to western blot analysis to detect full-length and cleaved Caspase-3 and PARP.

    Article Snippet: The cleavage of PARP and caspase-3 was monitored using anti-PARP (cat#95425), anti-caspase3 (cat#96625) and anti-cleaved caspase-3 (cat#96615) (all from Cell Signaling Technology, Beverly, MA, USA).

    Techniques: Cell Viability Assay, Standard Deviation, Western Blot

    Histological analysis of LV and liver and LV apoptosis 5 weeks after UCn3 gene transfer. (A) Hematoxylin and eosin (H E) and Masson's trichrome staining were performed on liver and transmural sections of LV (original magnification 20 × ; presented image magnification H E and Masson's trichrome: 7 × ), no evidence of inflammatory infiltrates was seen. LV fibrosis of the viable (non-infarcted LV) was similar in all three groups 8 weeks after MI. There were no group differences in liver fibrosis. (B) LV apoptosis was assessed using active caspase-3 staining on transmural LV sections, which showed a group difference between control and HF groups. Standard, control for Masson's trichrome. Individual data are shown (mean ± SE); p -values are from a Student's t

    Journal: Human Gene Therapy

    Article Title: Urocortin 3 Gene Transfer Increases Function of the Failing Murine Heart

    doi: 10.1089/hum.2018.103

    Figure Lengend Snippet: Histological analysis of LV and liver and LV apoptosis 5 weeks after UCn3 gene transfer. (A) Hematoxylin and eosin (H E) and Masson's trichrome staining were performed on liver and transmural sections of LV (original magnification 20 × ; presented image magnification H E and Masson's trichrome: 7 × ), no evidence of inflammatory infiltrates was seen. LV fibrosis of the viable (non-infarcted LV) was similar in all three groups 8 weeks after MI. There were no group differences in liver fibrosis. (B) LV apoptosis was assessed using active caspase-3 staining on transmural LV sections, which showed a group difference between control and HF groups. Standard, control for Masson's trichrome. Individual data are shown (mean ± SE); p -values are from a Student's t

    Article Snippet: After quenching of endogenous peroxidase activity and blocking with normal goat serum, the tissue sections were incubated (18 h at 4°C) with anti-cleaved caspase-3 antibody (1:200; Cell Signaling Technology) to assess apoptosis.

    Techniques: Staining

    Cleaved caspase-3 expression in the ovaries from each group. (A) Western blots presenting the expression of cleaved caspase-3 in the different groups. (B) Quantification of cleaved caspase-3 protein expression normalized to β-actin. Data are expressed as the mean ± standard error of the mean. One-way analysis of variance was used to analyze the data. # P

    Journal: Experimental and Therapeutic Medicine

    Article Title: Effects of dimethyl carbonate-induced autophagic activation on follicular development in the mouse ovary

    doi: 10.3892/etm.2017.5328

    Figure Lengend Snippet: Cleaved caspase-3 expression in the ovaries from each group. (A) Western blots presenting the expression of cleaved caspase-3 in the different groups. (B) Quantification of cleaved caspase-3 protein expression normalized to β-actin. Data are expressed as the mean ± standard error of the mean. One-way analysis of variance was used to analyze the data. # P

    Article Snippet: Membranes were then incubated with anti-cleaved caspase-3 antibody (1:1,000; cat. no. 5A1E; Cell Signaling Technology, Danvers, MA, USA), anti-Bcl-2 antibody (1:1,000; cat. no. 12789; Proteintech Group, Wuhan, China), anti-Bax antibody (1:2,000; cat. no. 60,267; Proteintech Group), anti-LC-3II antibody (1:1,000; cat. no. ab48394; Abcam), anti-Beclin-1 antibody (1:1,500; cat. no. 11306; Proteintech Group), anti-p62 antibody (1:1,000, cat. no. ab56416; Abcam), anti-HIF-1α antibody (1:500, cat. no. sc-53,546; Santa Cruz Biotechnology), anti-BNIP3 antibody (1:1,000, cat. no. ab10433; Abcam) and anti-β-actin antibody (1:4,000; cat. no. 66,009; Proteintech Group) overnight at 4°C.

    Techniques: Expressing, Western Blot

    Compound C abolished melatonin-induced cardioprotective effect on myocardial ischemia/reperfusion injury in type 1 diabetic rats. Myocardial ischemia/reperfusion surgery was performed after 1 month of streptozotocin injection. ( a ) Left ventricular systolic pressure. ( b ) and ( c ) The first derivative of left ventricular pressure (+dP/dt max and −dP/dt max ). Cardiac functional data was continuously monitored during the ischemia (30 min) and reperfusion period (3 hours). ( d ) Myocardial infarct size. ( e ) In situ detection of apoptotic cardiomyocytes by TUNEL staining (200×). ( f ) Myocardial apoptotic index. ( g ) Representative blots. ( h ) caspase-3 expression. ( i ) Bcl-2 expression. ( j ) Bax expression. ( k ) Cleaved caspase-3 expression. The depicted data are the means ± SEM, n = 6/group. ## P

    Journal: Scientific Reports

    Article Title: Melatonin ameliorates myocardial ischemia/reperfusion injury in type 1 diabetic rats by preserving mitochondrial function: role of AMPK-PGC-1α-SIRT3 signaling

    doi: 10.1038/srep41337

    Figure Lengend Snippet: Compound C abolished melatonin-induced cardioprotective effect on myocardial ischemia/reperfusion injury in type 1 diabetic rats. Myocardial ischemia/reperfusion surgery was performed after 1 month of streptozotocin injection. ( a ) Left ventricular systolic pressure. ( b ) and ( c ) The first derivative of left ventricular pressure (+dP/dt max and −dP/dt max ). Cardiac functional data was continuously monitored during the ischemia (30 min) and reperfusion period (3 hours). ( d ) Myocardial infarct size. ( e ) In situ detection of apoptotic cardiomyocytes by TUNEL staining (200×). ( f ) Myocardial apoptotic index. ( g ) Representative blots. ( h ) caspase-3 expression. ( i ) Bcl-2 expression. ( j ) Bax expression. ( k ) Cleaved caspase-3 expression. The depicted data are the means ± SEM, n = 6/group. ## P

    Article Snippet: Then, they were transferred to polyvinylidene difluoride membrane (Millipore, USA) and incubated overnight (4 °C) with p-AMPK, AMPK, PGC-1α, Bcl-2, Bax, caspase-3 and cleaved caspase-3 antibodies (Cell Signaling Technology, MA, USA, 1:1000 dilution), SIRT3, SOD2, NRF1, TFAM, cytochrome c and β-actin antibodies (Santa Cruz, CA, USA, 1:500 dilution) and total OXPHOS antibody cocktail (Abcam biotechnology, Cambridge, UK, 1:1000 dilution).

    Techniques: Injection, Functional Assay, In Situ, TUNEL Assay, Staining, Expressing

    Compound C and SIRT3 siRNA transfection blunted melatonin-induced anti-apoptotic effect against SIR injury in high glucose medium treated H9c2 cells. The H9c2 cells were exposed to high glucose medium (33 mmol/l) for 6 hours before the SIR treatment and during the entire reperfusion period (4 hours). HG-treatment was co-administered with or without melatonin (10 μmol/l) to evaluate its cytoprotective effect. Compound C (3 μmol/l) was administered for 6 hours before the SIR exposure to inhibit the AMPK signaling. ( a ) Cellular viability was presented by dividing the optical density of samples with that of the HG group. ( b ) Representative images of TUNEL staining (200×). ( c ) Percentage of TUNEL positive nuclei. ( d ) Cellular morphology (200×). ( e ) Representative blots. ( f ) Caspase-3 expression. ( g ) Cleaved caspase-3 expression. The depicted data are the means ± SEM, n = 6/group. ρρ P

    Journal: Scientific Reports

    Article Title: Melatonin ameliorates myocardial ischemia/reperfusion injury in type 1 diabetic rats by preserving mitochondrial function: role of AMPK-PGC-1α-SIRT3 signaling

    doi: 10.1038/srep41337

    Figure Lengend Snippet: Compound C and SIRT3 siRNA transfection blunted melatonin-induced anti-apoptotic effect against SIR injury in high glucose medium treated H9c2 cells. The H9c2 cells were exposed to high glucose medium (33 mmol/l) for 6 hours before the SIR treatment and during the entire reperfusion period (4 hours). HG-treatment was co-administered with or without melatonin (10 μmol/l) to evaluate its cytoprotective effect. Compound C (3 μmol/l) was administered for 6 hours before the SIR exposure to inhibit the AMPK signaling. ( a ) Cellular viability was presented by dividing the optical density of samples with that of the HG group. ( b ) Representative images of TUNEL staining (200×). ( c ) Percentage of TUNEL positive nuclei. ( d ) Cellular morphology (200×). ( e ) Representative blots. ( f ) Caspase-3 expression. ( g ) Cleaved caspase-3 expression. The depicted data are the means ± SEM, n = 6/group. ρρ P

    Article Snippet: Then, they were transferred to polyvinylidene difluoride membrane (Millipore, USA) and incubated overnight (4 °C) with p-AMPK, AMPK, PGC-1α, Bcl-2, Bax, caspase-3 and cleaved caspase-3 antibodies (Cell Signaling Technology, MA, USA, 1:1000 dilution), SIRT3, SOD2, NRF1, TFAM, cytochrome c and β-actin antibodies (Santa Cruz, CA, USA, 1:500 dilution) and total OXPHOS antibody cocktail (Abcam biotechnology, Cambridge, UK, 1:1000 dilution).

    Techniques: Transfection, TUNEL Assay, Staining, Expressing

    Western Blot with analysis per time point of WT and TLR−/− showing median and interquartile ranges with statistically significant differences at chronic time points for GAP43 and CASP3, as well as for synaptic markers of DLG4 and synaptophysin. ***,** and *marking P

    Journal: Scientific Reports

    Article Title: Tlr2 Deficiency is Associated with Enhanced Elements of Neuronal Repair and Caspase 3 Activation Following Brain Ischemia

    doi: 10.1038/s41598-019-39541-3

    Figure Lengend Snippet: Western Blot with analysis per time point of WT and TLR−/− showing median and interquartile ranges with statistically significant differences at chronic time points for GAP43 and CASP3, as well as for synaptic markers of DLG4 and synaptophysin. ***,** and *marking P

    Article Snippet: Primary antibodies were β3-Tubulin (5568 S, Cell Signalling Technology, Beverly, MA, USA) as a loading control, DLG4 (PSD95) (3450, Cell Signalling Technology Beverly, MA, USA), GAP43 (AB5220, Merck, Kenilworth, NJ, USA), synaptophysin clone SY38 (MAB5258, Merck, Kenilworth, NJ, USA), and anti-cleaved CASP3 (Cell Signaling, Danvers, MA, USA).

    Techniques: Western Blot

    Premature apoptosis occured in C9ORF72 spinal MNs over ageing (a) Apoptosis in MAP2-positive neurons (in red) was revealed as cytosolic rim staining around Hoechst-positive nuclei (in blue) for cleaved caspase 3 (Casp3, in green) by confocal IF microscopy at D80 endpoints (fig. 3). Dotted boxed areas in image galleries are shown magnified on the right. Note the striking accumulation of Casp3 in parental C9 and C9-KO that was phenocopied by WT-KO. Conversely, apoptosis hardly occurred in C9-GC and parental Ctrl1. Scale bars = 10µm. (b) Quantification of (a), percentage of apoptotic cells in MAP2-positive population. Asterisks: highly significant increase in any pairwise comparison with unlabeled conditions, one-way ANOVA with Bonferroni post test, *P≤0.05, **P≤0.01, ***P≤0.001, N=60 images from 3 independent experiments, error bars=SD. All unlabeled conditions (i.e. with no asterisk) were not significantly different amongst themselves in any pairwise comparison. Remaining comparisons were: WT-KO vs C9: ***, WT-KO vs C9-KO: ***, C9 vs C9-KO: ***.

    Journal: bioRxiv

    Article Title: High content live profiling reveals concomitant gain and loss of function pathomechanisms in C9ORF72 amyotrophic lateral sclerosis

    doi: 10.1101/2020.04.15.040394

    Figure Lengend Snippet: Premature apoptosis occured in C9ORF72 spinal MNs over ageing (a) Apoptosis in MAP2-positive neurons (in red) was revealed as cytosolic rim staining around Hoechst-positive nuclei (in blue) for cleaved caspase 3 (Casp3, in green) by confocal IF microscopy at D80 endpoints (fig. 3). Dotted boxed areas in image galleries are shown magnified on the right. Note the striking accumulation of Casp3 in parental C9 and C9-KO that was phenocopied by WT-KO. Conversely, apoptosis hardly occurred in C9-GC and parental Ctrl1. Scale bars = 10µm. (b) Quantification of (a), percentage of apoptotic cells in MAP2-positive population. Asterisks: highly significant increase in any pairwise comparison with unlabeled conditions, one-way ANOVA with Bonferroni post test, *P≤0.05, **P≤0.01, ***P≤0.001, N=60 images from 3 independent experiments, error bars=SD. All unlabeled conditions (i.e. with no asterisk) were not significantly different amongst themselves in any pairwise comparison. Remaining comparisons were: WT-KO vs C9: ***, WT-KO vs C9-KO: ***, C9 vs C9-KO: ***.

    Article Snippet: The following primary antibodies were used: mouse anti-yH2A.X (1:500, Millipore #05- 636), rabbit anti-53BP1 (1:1000, Novusbio NB100-304), chicken anti-MAP2 (1:1000, Abcam ab5392), rabbit anti-Casp3 (1:1000, Cell Signaling #9661), rat anti-GP (1:500, clone 18H8) and mouse anti GA (1:500, clone IAI2) were both generously provided from Dieter Edbauer ( ).

    Techniques: Staining, Microscopy

    Adeno-associated virus (AAV)-AEG-1 transduction of dopaminergic (DA) neurons in the in vivo SN of healthy mice. a Experimental schematic and the immunostaining for green fluorescent protein (GFP; green) and hemagglutinin (HA; brown) in the SNpc, which is outlined by the dotted elliptical shape, which was conducted following each viral injection. Scale bar, 200 μm. b Representative double immunofluorescent labeling of TH (red) and GFP (green) or TH and HA (green) in the SNpc. Scale bar, 20 μm. c Representative double immunofluorescent labeling for glial fibrillary acidic protein (GFAP)/ionized calcium binding adaptor molecule 1 (Iba1; red), which are markers of astrocytes and microglia, respectively, and GFP/HA (green) in the SNpc of healthy mice. Scale bar, 20 μm. d Immunostaining for TH in the SN and striatum (STR). Scale bars, 200 μm (black) and 50 μm (white) for the SN, and 1000 μm for the STR. e , f The number and optical density of the nigral TH-positive neurons and striatal TH-positive fibers, respectively (one-way ANOVA with Tukey’s post hoc test; n = 4 for each group). CON contralateral side, IPSI ipsilateral side. g Western blot analyses of the levels of cleaved caspase-3 (c-caspase-3) and cleaved poly (ADP-ribose) polymerase 1 (c-PARP-1) following AEG-1 transduction in the SN of healthy brains. * p = 0.005 vs . CON (one-way ANOVA with Tukey’s post hoc test; n = 4 for each group)

    Journal: Cell Death & Disease

    Article Title: Upregulation of neuronal astrocyte elevated gene-1 protects nigral dopaminergic neurons in vivo

    doi: 10.1038/s41419-018-0491-3

    Figure Lengend Snippet: Adeno-associated virus (AAV)-AEG-1 transduction of dopaminergic (DA) neurons in the in vivo SN of healthy mice. a Experimental schematic and the immunostaining for green fluorescent protein (GFP; green) and hemagglutinin (HA; brown) in the SNpc, which is outlined by the dotted elliptical shape, which was conducted following each viral injection. Scale bar, 200 μm. b Representative double immunofluorescent labeling of TH (red) and GFP (green) or TH and HA (green) in the SNpc. Scale bar, 20 μm. c Representative double immunofluorescent labeling for glial fibrillary acidic protein (GFAP)/ionized calcium binding adaptor molecule 1 (Iba1; red), which are markers of astrocytes and microglia, respectively, and GFP/HA (green) in the SNpc of healthy mice. Scale bar, 20 μm. d Immunostaining for TH in the SN and striatum (STR). Scale bars, 200 μm (black) and 50 μm (white) for the SN, and 1000 μm for the STR. e , f The number and optical density of the nigral TH-positive neurons and striatal TH-positive fibers, respectively (one-way ANOVA with Tukey’s post hoc test; n = 4 for each group). CON contralateral side, IPSI ipsilateral side. g Western blot analyses of the levels of cleaved caspase-3 (c-caspase-3) and cleaved poly (ADP-ribose) polymerase 1 (c-PARP-1) following AEG-1 transduction in the SN of healthy brains. * p = 0.005 vs . CON (one-way ANOVA with Tukey’s post hoc test; n = 4 for each group)

    Article Snippet: Materials Materials were purchased from the following companies: 6-OHDA (Sigma, St Louis, MO), desipramine (Sigma), l -ascorbic acid (Sigma), rabbit anti-TH (Pel-Freez, Brown Deer, WI), mouse anti-TH (R & D Systems, Minneapolis, MN), rabbit anti-Iba1 (Wako Pure Chemical Industries, Osaka, Japan), rabbit anti-GFAP (Millipore, Billerica, MA), rabbit anti-AEG-1 (Invitrogen, Camarillo, CA), rabbit anti-GFP (Millipore), mouse anti-HA (Cell Signaling, Beverly, MA), rabbit anti-HA (Cell Signaling), rabbit anti-FLAG (Sigma), rabbit anti-caspase-3 (Cell Signaling), rabbit anti-cleaved caspase-3 (Cell Signaling), rabbit anti-PARP-1 (Cell Signaling), rabbit anti-cleaved PARP-1 (Cell Signaling), rabbit anti-LC3B (Cell Signaling), rabbit anti-4E-BP1 (Cell Signaling), rabbit anti-p-4E-BP1 (Cell Signaling), mouse anti-NeuN (Millipore), rabbit anti-Akt (Cell Signaling), rabbit anti-p-Akt (Cell Signaling), rabbit anti-β-actin (Cell Signaling), rabbit anti-α-tubulin (Cell Signaling), mouse anti-Bcl-2 (Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-Bax (Santa Cruz Biotechnology), rabbit anti-p62/SQSTM1 (Sigma), biotinylated anti-rabbit IgG (Vector laboratories, Burlingame, CA), Texas Red-conjugated anti-rabbit/mouse IgG (Vector Laboratories), fluorescein (FITC)-conjugated anti-mouse IgG (Vector Laboratories), FITC-conjugated anti-rabbit IgG (Jackson ImmunoResearch Laboratories, Bar Harbor, ME), horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (Enzo Life Sciences, Farmingdale, NY) and HRP-conjugated anti-mouse IgG (Thermo Fisher Scientific, Rockford, IL).

    Techniques: Transduction, In Vivo, Mouse Assay, Immunostaining, Injection, Labeling, Binding Assay, Western Blot

    Anti-apoptotic effects of AEG-1 transduction in DA neurons on 6-OHDA neurotoxicity. a Western blot analyses show a significant increase in the levels of caspase-3, c-caspase-3, and c-PARP1 in the postmortem tissues of patients with PD compared with CON. p = 0.014 for caspase-3, p = 0.019 for c-caspase-3, and p = 0.009 for c-PARP-1, vs . CON ( t -test; n = 4 for each group). b Experimental schematic for Fig. 3c, d. c Representative double immunofluorescence labeling for TH (red) and c-caspase-3 (green) or TH and c-PARP-1 (green) in the mouse SN. AEG-1 upregulation induces reductions in the levels of expression of both c-caspase-3 and c-PARP-1 in TH-positive DA neurons in the SN with 6-OHDA neurotoxicity. Scale bar, 20 μm. d Western blot analyses of the levels of c-caspase-3 and c-PARP-1 in the SN 2 days after 6-OHDA treatment. * p = 0.029, ** p = 0.025, # p = 0.032, and ## p = 0.016 vs . CON; *** p = 0.009 and ### p = 0.002 vs . 6-OHDA alone (one-way ANOVA with Tukey’s post hoc test; n = 4 for each group)

    Journal: Cell Death & Disease

    Article Title: Upregulation of neuronal astrocyte elevated gene-1 protects nigral dopaminergic neurons in vivo

    doi: 10.1038/s41419-018-0491-3

    Figure Lengend Snippet: Anti-apoptotic effects of AEG-1 transduction in DA neurons on 6-OHDA neurotoxicity. a Western blot analyses show a significant increase in the levels of caspase-3, c-caspase-3, and c-PARP1 in the postmortem tissues of patients with PD compared with CON. p = 0.014 for caspase-3, p = 0.019 for c-caspase-3, and p = 0.009 for c-PARP-1, vs . CON ( t -test; n = 4 for each group). b Experimental schematic for Fig. 3c, d. c Representative double immunofluorescence labeling for TH (red) and c-caspase-3 (green) or TH and c-PARP-1 (green) in the mouse SN. AEG-1 upregulation induces reductions in the levels of expression of both c-caspase-3 and c-PARP-1 in TH-positive DA neurons in the SN with 6-OHDA neurotoxicity. Scale bar, 20 μm. d Western blot analyses of the levels of c-caspase-3 and c-PARP-1 in the SN 2 days after 6-OHDA treatment. * p = 0.029, ** p = 0.025, # p = 0.032, and ## p = 0.016 vs . CON; *** p = 0.009 and ### p = 0.002 vs . 6-OHDA alone (one-way ANOVA with Tukey’s post hoc test; n = 4 for each group)

    Article Snippet: Materials Materials were purchased from the following companies: 6-OHDA (Sigma, St Louis, MO), desipramine (Sigma), l -ascorbic acid (Sigma), rabbit anti-TH (Pel-Freez, Brown Deer, WI), mouse anti-TH (R & D Systems, Minneapolis, MN), rabbit anti-Iba1 (Wako Pure Chemical Industries, Osaka, Japan), rabbit anti-GFAP (Millipore, Billerica, MA), rabbit anti-AEG-1 (Invitrogen, Camarillo, CA), rabbit anti-GFP (Millipore), mouse anti-HA (Cell Signaling, Beverly, MA), rabbit anti-HA (Cell Signaling), rabbit anti-FLAG (Sigma), rabbit anti-caspase-3 (Cell Signaling), rabbit anti-cleaved caspase-3 (Cell Signaling), rabbit anti-PARP-1 (Cell Signaling), rabbit anti-cleaved PARP-1 (Cell Signaling), rabbit anti-LC3B (Cell Signaling), rabbit anti-4E-BP1 (Cell Signaling), rabbit anti-p-4E-BP1 (Cell Signaling), mouse anti-NeuN (Millipore), rabbit anti-Akt (Cell Signaling), rabbit anti-p-Akt (Cell Signaling), rabbit anti-β-actin (Cell Signaling), rabbit anti-α-tubulin (Cell Signaling), mouse anti-Bcl-2 (Santa Cruz Biotechnology, Santa Cruz, CA), mouse anti-Bax (Santa Cruz Biotechnology), rabbit anti-p62/SQSTM1 (Sigma), biotinylated anti-rabbit IgG (Vector laboratories, Burlingame, CA), Texas Red-conjugated anti-rabbit/mouse IgG (Vector Laboratories), fluorescein (FITC)-conjugated anti-mouse IgG (Vector Laboratories), FITC-conjugated anti-rabbit IgG (Jackson ImmunoResearch Laboratories, Bar Harbor, ME), horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (Enzo Life Sciences, Farmingdale, NY) and HRP-conjugated anti-mouse IgG (Thermo Fisher Scientific, Rockford, IL).

    Techniques: Transduction, Western Blot, Immunofluorescence, Labeling, Expressing

    Apoptosis of MIN6 cells induced by TM is inhibited by overexpression of DHCR24. (a) A representative fluorescent image of cleaved caspase-3 (green) was obtained by IC analysis 24 h after TM exposure. Scale bar, 50 μ m. (b) The relative fluorescence intensity of cleaved caspase-3 was analyzed with ImageJ software. Mean ± S.D. ( n = 3); ∗ p

    Journal: Journal of Diabetes Research

    Article Title: 3β-Hydroxysteroid-Δ24 Reductase (DHCR24) Protects Pancreatic β Cells from Endoplasmic Reticulum Stress-Induced Apoptosis by Scavenging Excessive Intracellular Reactive Oxygen Species

    doi: 10.1155/2020/3426902

    Figure Lengend Snippet: Apoptosis of MIN6 cells induced by TM is inhibited by overexpression of DHCR24. (a) A representative fluorescent image of cleaved caspase-3 (green) was obtained by IC analysis 24 h after TM exposure. Scale bar, 50 μ m. (b) The relative fluorescence intensity of cleaved caspase-3 was analyzed with ImageJ software. Mean ± S.D. ( n = 3); ∗ p

    Article Snippet: Briefly, cells on coverslips were fixed and blocked, followed by incubation with a mouse anti-myc antibody (Santa Cruz Biotechnology, CA, USA) or rabbit anti-cleaved caspase-3 antibody (Cell Signaling, Beverly, MA) overnight at 4°C.

    Techniques: Over Expression, Fluorescence, Software

    Quantitative analysis of cleaved caspase-3 + cells per high power field (HPF) ( a ) and of BrdU + hepatocytes per HPF ( b ) in liver tissue of sham-treated, of combination (combi)-treated as well as of mono (HPβCD)-treated Npc1 +/+ (sham, n = 7 (BrdU n = 3); combi, n = 9 (BrdU n = 3); mono, n = 15 (BrdU n = 3) and Npc1 −/− mice (sham, n = 10 (BrdU n = 3); combi, n = 10 (BrdU n = 3); mono, n = 11 (BrdU n = 3)) with representative light microscopic images of liver tissue of sham-treated, combi-treated and mono-treated Npc1 −/− mice. Note the significant reduction of apoptosis and proliferation in combi-treated Npc1 −/− mice. Values are given as mean ± SEM; ANOVA; post-hoc pairwise comparison tests: * p

    Journal: International Journal of Molecular Sciences

    Article Title: Evaluation of Two Liver Treatment Strategies in a Mouse Model of Niemann–Pick-Disease Type C1

    doi: 10.3390/ijms19040972

    Figure Lengend Snippet: Quantitative analysis of cleaved caspase-3 + cells per high power field (HPF) ( a ) and of BrdU + hepatocytes per HPF ( b ) in liver tissue of sham-treated, of combination (combi)-treated as well as of mono (HPβCD)-treated Npc1 +/+ (sham, n = 7 (BrdU n = 3); combi, n = 9 (BrdU n = 3); mono, n = 15 (BrdU n = 3) and Npc1 −/− mice (sham, n = 10 (BrdU n = 3); combi, n = 10 (BrdU n = 3); mono, n = 11 (BrdU n = 3)) with representative light microscopic images of liver tissue of sham-treated, combi-treated and mono-treated Npc1 −/− mice. Note the significant reduction of apoptosis and proliferation in combi-treated Npc1 −/− mice. Values are given as mean ± SEM; ANOVA; post-hoc pairwise comparison tests: * p

    Article Snippet: For immunohistochemical analysis of cleaved caspase-3+ cells, F4/80+ von Kupffer cells (resident liver macrophages) and BrdU+ hepatocytes sections were pre-treated with citrate puffer (pH 6.0) in the microwave (700 W for 7 min) and were either exposed to a rabbit anti-cleaved caspase-3 antibody (1:500, Cell Signaling Technology, Frankfurt, Germany), a rat anti-F4/80 (1:10; Serotec, Oxford, UK) or a mouse anti-BrdU antiserum (1:50; DakoCytomation, Dako, Hamburg, Germany).

    Techniques: Mouse Assay

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

    Journal: Frontiers in Neurology

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

    doi: 10.3389/fneur.2018.00931

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

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

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

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

    Journal: Frontiers in Neurology

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

    doi: 10.3389/fneur.2018.00931

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

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

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

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

    Journal: Chinese Medical Journal

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

    doi: 10.4103/0366-6999.245158

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

    Article Snippet: The primary antibodies (1:1000 diluted in Tris-buffered saline with 0.1% tween, TBST) were used to detect COQ6, nephrin, cleaved caspase-3 (Abcam, MA, USA), F-actin (Abcam, MA, USA), and β-actin (Abcam, MA, USA). β-actin was used as an internal control.

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

    Effects of CLU silencing on viability, proliferation/death and differentiation of cultured OA chondrocytes. ( A ) CLU mRNA and protein expression after 48 and 72 hours, respectively, in scramble (control) and siCLU transfected OA-HACs. ( B ) Cell viability by MTT assay after 24, 48 and 72 hours of scramble and siCLU transfected OA-HACs. ( C ) Representative immunocytochemical images and bar graphs showing Ki67 + and p21 + nuclei in cultured OA-HACs and ( D ) cleaved caspase-3 + cytospinned OA-HACs after 72 hours of silencing. ( E ) Gene array shows the modulation of NFkB and Stat-3-related genes in siCLU transfected OA-HACs compared with scramble after 48 hours of silencing. Transcript levels were normalized on GAPDH (internal reference gene). ( F ) mRNA levels by Real-Time PCR of COL2A1, ACAN, MMP13, COL10A1, and ( G ) representative blot and densitometric analysis of MMP13 and COL10A1 in scramble and CLU-silenced OA-HACs. Data are expressed as mean values ± SEM of three independent experiments (each using pooled samples from n=3 OA donors) performed in triplicate. Abbreviations: OA-HACs, OA-Human articular chondrocytes; OD, optical density unit; RLU, relative light unit. Student’s t-test: * P

    Journal: Aging (Albany NY)

    Article Title: Clusterin exerts a cytoprotective and antioxidant effect in human osteoarthritic cartilage

    doi: 10.18632/aging.103310

    Figure Lengend Snippet: Effects of CLU silencing on viability, proliferation/death and differentiation of cultured OA chondrocytes. ( A ) CLU mRNA and protein expression after 48 and 72 hours, respectively, in scramble (control) and siCLU transfected OA-HACs. ( B ) Cell viability by MTT assay after 24, 48 and 72 hours of scramble and siCLU transfected OA-HACs. ( C ) Representative immunocytochemical images and bar graphs showing Ki67 + and p21 + nuclei in cultured OA-HACs and ( D ) cleaved caspase-3 + cytospinned OA-HACs after 72 hours of silencing. ( E ) Gene array shows the modulation of NFkB and Stat-3-related genes in siCLU transfected OA-HACs compared with scramble after 48 hours of silencing. Transcript levels were normalized on GAPDH (internal reference gene). ( F ) mRNA levels by Real-Time PCR of COL2A1, ACAN, MMP13, COL10A1, and ( G ) representative blot and densitometric analysis of MMP13 and COL10A1 in scramble and CLU-silenced OA-HACs. Data are expressed as mean values ± SEM of three independent experiments (each using pooled samples from n=3 OA donors) performed in triplicate. Abbreviations: OA-HACs, OA-Human articular chondrocytes; OD, optical density unit; RLU, relative light unit. Student’s t-test: * P

    Article Snippet: After permeabilization in TBS-0.1% Tween 20 for 5min at RT, slides were incubated with rabbit polyclonal antibody anti-Ki67 (Novus Biologicals, CO, USA, 1:100, 1h at RT), goat polyclonal anti-CLU β (Santa Cruz Biotechnology, 1:50, O/N at +4°C), mouse monoclonal anti-p21 (R & D System Inc., USA, 8μg/ml, 1h at RT), rabbit polyclonal anti-cleaved caspase-3 (Cell Signaling Technology, Inc, MA, USA, 1:500, 1h at RT) and goat polyclonal anti-TNFα (Santa Cruz Biotechnology, 1:100, 2h at RT), with positive and negative controls.

    Techniques: Cell Culture, Expressing, Transfection, MTT Assay, Real-time Polymerase Chain Reaction

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

    Journal: Oncotarget

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

    doi: 10.18632/oncotarget.22776

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

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

    Techniques: Expressing, Western Blot

    Protective effect of TUCDA against ER stress-induced apoptosis of MSCs. ( a ) Western blot analysis showing the expression of BCL-2, Bax, cleaved caspase-3, and cleaved PARP-1 after treatment of MSCs with H 2 O 2 (200 μM) for the indicated times (0, 2, 4, 6, or 8 h). ( b ) The expression levels of BCL-2, Bax, cleaved caspase-3, and cleaved PARP-1 were normalized to that of β-actin. Values represent the mean ± SEM. ** P

    Journal: Scientific Reports

    Article Title: Tauroursodeoxycholic acid reduces ER stress by regulating of Akt-dependent cellular prion protein

    doi: 10.1038/srep39838

    Figure Lengend Snippet: Protective effect of TUCDA against ER stress-induced apoptosis of MSCs. ( a ) Western blot analysis showing the expression of BCL-2, Bax, cleaved caspase-3, and cleaved PARP-1 after treatment of MSCs with H 2 O 2 (200 μM) for the indicated times (0, 2, 4, 6, or 8 h). ( b ) The expression levels of BCL-2, Bax, cleaved caspase-3, and cleaved PARP-1 were normalized to that of β-actin. Values represent the mean ± SEM. ** P

    Article Snippet: Immunofluorescence staining was performed using primary antibodies against human nuclear antigen (HNA; Millipore, Billerica, MA, USA), manganese-dependent superoxide dismutase (MnSOD; Santa Cruz Biotechnology, Santa Cruz, CA, USA), cleaved caspase-3 (Santa Cruz Biotechnology), CD31 (Santa Cruz Biotechnology), and α-SMA (Santa Cruz Biotechnology) and secondary antibodies Alexa-488 and Alexa-594 (Thermo Fisher Scientific, Waltham, MA, USA).

    Techniques: Western Blot, Expressing

    Screening OB2C small molecule library for death ligands Cleaved caspase 3 positive beads (red arrow) in OB2C beads library. Positive beads were picked up for chemical decoding with automatic Edman microsequencing. ( a ) LLS1, ( b ) LLS2. Resynthesize death ligand LLS2 on beads for validation of death effect. ( c ) Cleaved caspase 3 staining.(100X) ( d ) Propidium Iodide staining for dead cells. (40X). Galectin-1 is a target protein of LLS2. (e) Eluted protein from pull-down assay was identified as galectin-1 by LC MS/MS. (f) Immunoblot analysis with anti-galectin-1 Ab, to validate the LC MS/MS result. 1: blank bead, 2: LLS2-beasd, 3: a irrelevant small molecule-bead control. (g) Immunostaining result reveals that LLS2 co-localizes with galectin-1. SKOV3, A549, PC3 and XPA3 cells were stained by biotinylated LLS2 (green), galectin-1 antibody (red), and nuclei were stained with DAPI (blue). (h) Computer modeling shows that LLS2 binds the interface of the galectin-1 dimer. Residues that are within 3.5 angstroms from LLS2 were shown. Scale bars: 50 μm.

    Journal: Molecular cancer therapeutics

    Article Title: A Novel Galectin-1 Inhibitor Discovered through One-Bead-Two-Compounds Library Potentiates the Anti-tumor Effects of Paclitaxel in vivo

    doi: 10.1158/1535-7163.MCT-16-0690

    Figure Lengend Snippet: Screening OB2C small molecule library for death ligands Cleaved caspase 3 positive beads (red arrow) in OB2C beads library. Positive beads were picked up for chemical decoding with automatic Edman microsequencing. ( a ) LLS1, ( b ) LLS2. Resynthesize death ligand LLS2 on beads for validation of death effect. ( c ) Cleaved caspase 3 staining.(100X) ( d ) Propidium Iodide staining for dead cells. (40X). Galectin-1 is a target protein of LLS2. (e) Eluted protein from pull-down assay was identified as galectin-1 by LC MS/MS. (f) Immunoblot analysis with anti-galectin-1 Ab, to validate the LC MS/MS result. 1: blank bead, 2: LLS2-beasd, 3: a irrelevant small molecule-bead control. (g) Immunostaining result reveals that LLS2 co-localizes with galectin-1. SKOV3, A549, PC3 and XPA3 cells were stained by biotinylated LLS2 (green), galectin-1 antibody (red), and nuclei were stained with DAPI (blue). (h) Computer modeling shows that LLS2 binds the interface of the galectin-1 dimer. Residues that are within 3.5 angstroms from LLS2 were shown. Scale bars: 50 μm.

    Article Snippet: The tissue sections were then incubated with the specific antibody against cleaved-caspase 3 and Ki-67(Cell signaling) overnight, rinsed extensively three times with PBS.

    Techniques: Staining, Pull Down Assay, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Immunostaining

    LLS2 alone and LLS2/PTX have anti-tumor activity in SKOV3 xenograft model (a) Xenograft tumor. (b) Tumor growth curves, and (c) tumor weight of the xenografts in inoculated nude mice. (d) Body weight of nude mice. Briefly, 2.5 × 106 SKOV3 cells were subcutaneously injected to the right side of the dorsal flank of the female congenital athymic BALB/c nude mice. The tumors were allowed to grow to about 100 mm3. Then, mice were randomly divided into control and treatment groups (n=5). Mice were given a daily I.V. administartion for 5 successive days. (e) IHC detection of ki-67 and cleaved caspase-3 expression. (f) Quantification of immunostaining of ki-67 and cleaved caspase 3 positive cells. The cells were counted in 3 random chosen areas. * P

    Journal: Molecular cancer therapeutics

    Article Title: A Novel Galectin-1 Inhibitor Discovered through One-Bead-Two-Compounds Library Potentiates the Anti-tumor Effects of Paclitaxel in vivo

    doi: 10.1158/1535-7163.MCT-16-0690

    Figure Lengend Snippet: LLS2 alone and LLS2/PTX have anti-tumor activity in SKOV3 xenograft model (a) Xenograft tumor. (b) Tumor growth curves, and (c) tumor weight of the xenografts in inoculated nude mice. (d) Body weight of nude mice. Briefly, 2.5 × 106 SKOV3 cells were subcutaneously injected to the right side of the dorsal flank of the female congenital athymic BALB/c nude mice. The tumors were allowed to grow to about 100 mm3. Then, mice were randomly divided into control and treatment groups (n=5). Mice were given a daily I.V. administartion for 5 successive days. (e) IHC detection of ki-67 and cleaved caspase-3 expression. (f) Quantification of immunostaining of ki-67 and cleaved caspase 3 positive cells. The cells were counted in 3 random chosen areas. * P

    Article Snippet: The tissue sections were then incubated with the specific antibody against cleaved-caspase 3 and Ki-67(Cell signaling) overnight, rinsed extensively three times with PBS.

    Techniques: Activity Assay, Mouse Assay, Injection, Immunohistochemistry, Expressing, Immunostaining