Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Sepsis Induces Extensive Autophagic Vacuolization in Hepatocytes –a clinical and laboratory based study

doi: 10.1038/labinvest.2009.8

Figure Lengend Snippet: Apoptosis, autophagy and inflammation pathway genes evaluated in the present study

Article Snippet: Casp8 , caspase 8 , GO:0008624: induction of apoptosis by extracellular signals , NM_009812.2 , Mm00802247_m1.

Techniques: Transformation Assay, Binding Assay, Transduction, Full Display Name, Activation Assay

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Sepsis Induces Extensive Autophagic Vacuolization in Hepatocytes –a clinical and laboratory based study

doi: 10.1038/labinvest.2009.8

Figure Lengend Snippet: Apoptosis, autophagy and inflammation pathway gene expression on cecal ligation and puncture in total liver cells

Article Snippet: Casp8 , caspase 8 , GO:0008624: induction of apoptosis by extracellular signals , NM_009812.2 , Mm00802247_m1.

Techniques: Gene Expression, Ligation

Journal: The Journal of Biological Chemistry

Article Title: The long non-coding RNA HOTAIR enhances pancreatic cancer resistance to TNF-related apoptosis-inducing ligand

doi: 10.1074/jbc.M117.786830

Figure Lengend Snippet: Overexpression of HOTAIR attenuates TRA-8-induced apoptosis in sensitive pancreatic cancer cells. A and B, BxPC3 (A) and MiaPaCa-2 (B) cells were infected with lentiviruses carrying control vector or HOTAIR cDNA (HOTAIR), and stable clones were selected by puromycin. Panels Aa and Ba, HOTAIR expression, as determined by qRT-PCR and normalized by β-actin expression (n = 3, ***, p < 0.001). Panels Ab and Bb, TRA-8-induced apoptosis. BxPC3 and MiaPaCa-2 cells with HOTAIR overexpression and their control vector cells were seeded into 6-well plates at 2 × 105 per well. After culturing for 24 h, cells were exposed to TRA-8 (1 μg/ml) for 24 h, and apoptosis was determined by flow cytometry using Annexin PE and 7 AAD staining kit. TRA-8-induced apoptosis is shown in the hatched bars (n = 3, ***, p < 0.001). Panels Ac and Bc, Western blot analysis of the expression of caspase-8 (Casp8). The expression of β-actin was used as a loading control. Representative blots from three independent experiments are shown.

Article Snippet: Antibodies were purchased as follows: rabbit anti-DR5 antibody (ProSci, no. 2019, lot number 5355–1502), mouse anti-caspase 8 (BIOSOURCE no. AHZ0502, lot number 22363–01S), mouse anti-β-actin (Sigma, no. A5541–2MI, lot number 014M4759), rabbit anti-EZH2 (Cell Signaling Technology, no. D2C9, lot number 7), and mouse anti-trimethyl-histone H3 (lysine 27) (Active Motif, no. 61017, lot number 23115012).

Techniques: Over Expression, Infection, Plasmid Preparation, Clone Assay, Expressing, Quantitative RT-PCR, Flow Cytometry, Staining, Western Blot

Journal: Oncogene

Article Title: VPS34 stimulation of p62 phosphorylation for cancer progression

doi: 10.1038/onc.2017.295

Figure Lengend Snippet: Inhibition of caspase 8 promotes the fusion of autophagosome with lysosome. ( a ) Inhibition of LBH589-induced apoptosis by caspase inhibitors. Apoptotic cell death of MCF-7 cells induced by treatment with the indicated concentrations of LBH589 and/or caspase inhibitors for 48 h. For caspase inhibitor treatment, cells were pretreated with 10 μ M z-VAD-FMK (Z-VAD) or z-IETD-FMK (IETD) for 4 h. ( b ) Z-VAD or IETD inhibited LBH589-induced cleavage of caspase 7, caspase 8, caspase 9 and PARP-1 in MCF-7 cells treated as indicated for 24 h. ( c ) Cell viability of MCF-7 cells treated as indicated for 24 h. ( d ) Accumulation of Vps34, LC3 and p62 in MCF-7 cells treated as indicated with or without 20 μM chloroquine for 24 h. ( e ) Confocal microscopic evaluation of autophagosome-lysosome fusion in MCF-7 cells. MCF-7 cells stably expressing EGFP-LC3 (Green) treated as indicated for 18 h were stained with lysosome tracker staining (Red). Immunofluorescence analyses were performed using confocal microscopic detection (63 × oil). Left, representative confocal images. Right, relative Green + red + dots per cell calculated from 20 cells; bars, s.d. ** P <0.01. ( f ) Disruption of autophagosome-lysosome fusion in MCF-7 cells. MCF-7 cells stably expressing tfLC3 were treated as indicated for 18 h. Immunofluorescence analyses were performed using confocal microscopic detection (60 × oil). Left, representative confocal images. Right, % of autolysosomes per cell was calculated from 20 cells by using the formula of (1-yellow dots/red dots) × 100; bars, s.d. ** P <0.01. DAPI, 4', 6-diamidino-2-phenylindole.

Article Snippet: The other constructs were obtained as follows: pcDNA3-Casp8 (Addgene, 11817), pcDNA3-Casp8-Mt (Addgene, 11818).

Techniques: Inhibition, Stable Transfection, Expressing, Staining, Immunofluorescence, Disruption

Journal: Oncogene

Article Title: VPS34 stimulation of p62 phosphorylation for cancer progression

doi: 10.1038/onc.2017.295

Figure Lengend Snippet: Vps34 is cleaved by active caspase 8. ( a ) Active caspase 8 eliminated SUMOylated Vps34 and total Vps34. Immunoblot of His-SUMO1-conjugated Vps34 and total Vps34 in HEK293T cells transfected as indicated for 24 h, and treated with LBH589 for 24 h. ( b ) Except z-IETD-FMK (20 μ M ), neither MG132 (10 μ M ) nor chloroquine (20 μ M ) blocked abrogation of F-Vps34 caused by active caspase 8. Immunoblot analyses were performed on transfected HEK293T cells, as indicated. ( c ) Active caspase 8 degraded Vps34 in a dose-dependent manner. Vps34 was extracted from HEK293T cells transfected as indicated. ( d ) Immunostaining of endogenous Vps34 (green) colocalized with active caspase 8 (red) in MCF7 cells treated with LBH589 and/or z-IETD-FMK, for 18 h. ( e ) Abrogation of endogenous Vps34 cleavage by caspase inhibitors in LBH589-treated MCF-7 cells. The red arrow denotes cleaved Vps34. ( f ) Upper, immunoblot analyses of variant mutant of Vps34 expression in HEK293T cells with the presence of LBH589 for 24 h. Lower, the location of predicted cleavage sites in Vps34 structure of scheme. ( g ) Caspase 8 cleavage of Vps34 at the site of Asp285. Immunoblot of the variant mutant effects on Vps34 expression in HEK293T cells cotransfected with caspase 8 treated with 50 n M LBH589 for 24 h. ( h ) In vitro cleavage of Vps34 by active caspase 8. The red arrow denotes cleaved Vps34. ( i ) Multiple alignment of Vps34 shows conservation of the caspase 8 cleavage site in the SDHD motif. DAPI, 4', 6-diamidino-2-phenylindole.

Article Snippet: The other constructs were obtained as follows: pcDNA3-Casp8 (Addgene, 11817), pcDNA3-Casp8-Mt (Addgene, 11818).

Techniques: Western Blot, Transfection, Immunostaining, Variant Assay, Mutagenesis, Expressing, In Vitro

Journal: PLoS ONE

Article Title: Spatial and Temporal Gene Expression Differences in Core and Periinfarct Areas in Experimental Stroke: A Microarray Analysis

doi: 10.1371/journal.pone.0052121

Figure Lengend Snippet: Regulation of DNA replication, recombination & repair and also cell death-associated genes in the core and periinfarct after permanent focal cerebral ischemia.

Article Snippet: Real time PCR was performed using Taqman (Applied Biosystems) gene expression assays (GAPD, Rn01775763_g1; PGK1, Rn00821429_g1; HMBS, Rn00565886_m; BDNF, Rn02531967_s1; VEGFc, Rn01488076_m; OLIG2, Rn01767116_m; CASP8, Rn00574069_m; PCNA, Rn00574296_g1; LINGO3, Rn01408564_m; HSPA12a, Rn01410714_m; HES2, Rn00570311_g1; SEMA4f, Rn00570562_m; NEFM, Rn00566763_m; CXCL12, Rn00573260_m; CXCR4, Rn01483207_m; BMP4, Rn00432087_m; EGR1, Rn00561138_m; IGF1, Rn00710306_m; CALB1, Rn00583140_m; NRN1L, Rn01762396_g1; FGF18, Rn00433286_m; CXCL1, Rn00578225_m).

Techniques: