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tunel-tdt-mediated-dutp-nick-end-labeling

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ER stress-induced apoptosis of INS-1 cells. <em>A</em>: Suppression...
Published: 01 February 2004
FIG. 3. ER stress-induced apoptosis of INS-1 cells. A: Suppression of thapsigargin-induced apoptosis of iPLA2β-overexpressing INS-1 cells by BEL. INS-1 cells stably transfected with a vector containing iPLA2β cDNA (OE) construct were treated with thapsigargin (T) (1 μmol/l) for 24 h in the absence or presence of BEL (10 μmol/l). The cells were then harvested for TUNEL (Tdt-mediated dUTP nick end labeling) analyses to assess the magnitude of cell apoptosis (*T + BEL-treated group significantly different from T alone-treated group, P < 0.05). B: Immunoblotting analyses. Aliquots of INS-1 cell protein (50 μg), prepared from OE cells treated with dimethylsulfoxide (lane 1) or thapsigargin (1 μmol/l) (lane 2), were analyzed by SDS-PAGE (7.5%) and transferred onto an Immobolin-P polyvinylidene difluoride membrane. The electroblot was probed with piPLA2β antibodies, and immunoreactive protein bands were visualized by enhanced chemiluminescence, as described ( 41 ). C, control. C: Effects of caspase-3 inhibition on iPLA2β subcellular distribution. iPLA2β OE INS-1 cells seeded in glass chambered slides were pretreated with caspase-3 inhibitor (C3-I) (500 nmol/l) or vehicle for 24 h. The cells were then treated with thapsigargin in the absence and presence of C3-I for 24 h and subsequently processed for iPLA2β immunofluorescence analyses by confocal microscopy. Con, control. FIG. 3. ER stress-induced apoptosis of INS-1 cells. A: Suppression of thapsigargin-induced apoptosis of iPLA2β-overexpressing INS-1 cells by BEL. INS-1 cells stably transfected with a vector containing iPLA2β cDNA (OE) construct were treated with thapsigargin (T) (1 μmol/l) for 24 h in the absence or presence of BEL (10 μmol/l). The cells were then harvested for TUNEL (Tdt-mediated dUTP nick end labeling) analyses to assess the magnitude of cell apoptosis (*T + BEL-treated group significantly different from T alone-treated group, P < 0.05). B: Immunoblotting analyses. Aliquots of INS-1 cell protein (50 μg), prepared from OE cells treated with dimethylsulfoxide (lane 1) or thapsigargin (1 μmol/l) (lane 2), were analyzed by SDS-PAGE (7.5%) and transferred onto an Immobolin-P polyvinylidene difluoride membrane. The electroblot was probed with piPLA2β antibodies, and immunoreactive protein bands were visualized by enhanced chemiluminescence, as described (41). C, control. C: Effects of caspase-3 inhibition on iPLA2β subcellular distribution. iPLA2β OE INS-1 cells seeded in glass chambered slides were pretreated with caspase-3 inhibitor (C3-I) (500 nmol/l) or vehicle for 24 h. The cells were then treated with thapsigargin in the absence and presence of C3-I for 24 h and subsequently processed for iPLA2β immunofluorescence analyses by confocal microscopy. Con, control. More
Meeting Abstracts
Journal: Diabetes
Diabetes 2001;50(suppl_1):S48
Published: 01 February 2001
... cells are detected by the TdT-mediated dUTP nick-end labeling (TUNEL) method at a high frequency in both Kir6.2G372S Tg and Kir6.2-/- mice compared with the respective controls. Thus, studies of Kir6.2G372S Tg and Kir6.2-/- mice indicate that K(ATP) channels play an important role in cell survival...
Meeting Abstracts
Journal: Diabetes
Diabetes 1999;48(8):1652–1657
Published: 01 August 1999
... the focal lesion only. Using the Tdt-mediated dUTP nick end labeling (TUNEL) reaction, apoptotic beta-cells were identified exclusively within the focal region. At diagnosis, disease severity was similar in patients with paternally inherited mutations and those with two mutations. For patients who did...
Journal Articles
Journal: Diabetes
Diabetes 2010;59(7):1825–1835
Published: 27 April 2010
..., and CaMKII activity were all greatly increased in the retinas of diabetic mice compared with controls, 2 months after induction of diabetes. Terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL)-positive signals co-localized with CaMKII- and phospho-CaMKII immunoreactive RGCs...
Journal Articles
Journal: Diabetes
Diabetes 1997;46(7):1189–1197
Published: 01 July 1997
..., Joslin Diabetes Center, One Joslin Place, Boston, MA 02215. loekenm@joslab.harvard.edu . 1 NTD, neural tube defect; PBS, phosphate-buffered saline; RT-PCR, reverse transcription–polymerase chain reaction; SOD, superoxide dismutase; STZ, streptozotocin; TUNEL, Tdt-mediated dUTP nick end labeling...
Journal Articles
Journal: Diabetes
Diabetes 2003;52(7):1677–1682
Published: 01 July 2003
..., glucocorticoid receptor LOD, logarithm of odds NK, natural killer PI, propidium iodide QTL, quantitative trait locus TCR, T-cell receptor Dxm, dexamethazone TdT, terminal deoxynucleotidyl transferase Th, T helper TUNEL, TdT-mediated dUTP nick-end labeling The nonobese diabetic (NOD) mouse...
Journal Articles
Journal: Diabetes
Diabetes 2002;51(1):159–167
Published: 01 January 2002
.... For this reason, in the present study, the effects of AICAR on fatty-acid metabolism and on indexes of apoptosis, such as TdT-mediated dUTP nick-end labeling (TUNEL) staining and caspase-3 activity, were evaluated in HUVEC exposed to media enriched in glucose. In addition, the effect of expressing...
Journal Articles
Journal: Diabetes
Diabetes 2004;53(suppl_1):S179–S185
Published: 01 February 2004
... μmol/l) for 24 h in the absence or presence of BEL (10 μmol/l). The cells were then harvested for TUNEL (Tdt-mediated dUTP nick end labeling) analyses to assess the magnitude of cell apoptosis (*T + BEL-treated group significantly different from T alone-treated group, P < 0.05). B...
Journal Articles
Journal: Diabetes
Diabetes 2005;54(9):2541–2548
Published: 01 September 2005
...@ualberta.ca 20 6 2005 18 4 2005 DIABETES 2005 GAL, galactosidase Hu-XIAP, human XIAP TUNEL, TdT-mediated dUTP nick-end labeling XIAP, X-linked inhibitor of apoptosis protein The recent introduction of the Edmonton Protocol has demonstrated that islet transplantation...
Journal Articles
Journal: Diabetes
Diabetes 2004;53(8):2018–2023
Published: 01 August 2004
... factor PI, phosphatidylinositol TUNEL, Tdt-mediated dUTP nick-end labeling Inadequate β-cell mass is a crucial factor in diabetes. Immune destruction of β-cells is the main defect in type 1 diabetes ( 1 ), whereas in type 2 diabetes, β-cell mass is decreased to some extent compared with a normal...
Journal Articles
Journal: Diabetes
Diabetes 2005;54(1):212–219
Published: 01 January 2005
... 2004 7 7 2004 DIABETES 2005 AGE, advanced glycation end product FITC, fluorescein isothiocyanate NMDA, N-methyl-d-aspartate nNOS, neuronal nitric oxide synthase STZ, streptozotocin TUNEL, Tdt-mediated dUTP nick-end labeling VAChT, vesicular acetylcholine...
Journal Articles
Journal: Diabetes
Diabetes 2004;53(6):1509–1516
Published: 01 June 2004
... IAPP, islet amyloid polypeptide TUNEL, TdT-mediated dUTP nick-end labeling In humans with type 2 diabetes, insulin secretion is defective ( 1 , 2 ), presumably at least in part because of an ∼60% defect in β-cell mass when compared with BMI-matched nondiabetic humans ( 3 , 4 ). Several mechanisms...
Journal Articles
Journal: Diabetes
Diabetes 2005;54(6):1838–1845
Published: 01 June 2005
..., inhibitor of κB IKK, inhibitor of κB kinase NF-κB, nuclear factor κB PC, pericyte PEI, polyethylenimine TUNEL, TdT-mediated dUTP nick-end labeling Pericyte (PC) drop out due to apoptosis has long been considered an early event in diabetic retinopathy ( 1 ). Theories to explain why it occurs have...
Meeting Abstracts
Journal: Diabetes
Diabetes 1998;47(12):1948–1952
Published: 01 December 1998
... saline. Apo- 1948 DIABETES, VOL. 47, DECEMBER 1998 R. MAGUN AND ASSOCIATES ptosis was measured by TdT-mediated dUTP nick end labeling (TUNEL) using the ated adipocytes did not exhibit any positive staining for the In Situ Cell Death Detection kit (Boehringer Mannheim) according to manufac- same durations...
Meeting Abstracts
Journal: Diabetes
Diabetes 1999;48(12):2454–2462
Published: 01 December 1999
..., reverse transcription polymerase chain reaction; STZ, streptozotocin; TUNEL, Tdt-mediated dUTP nick end labeling. 2454 DIABETES, VOL. 48, DECEMBER 1999 E.L. FINE AND ASSOCIATES studies, including the Diabetes Control and Complications ture dishes that had been coated with 1% gelatin (Sigma) containing 2...
Journal Articles
Journal: Diabetes
Diabetes 2004;53(11):2804–2814
Published: 01 November 2004
..., proinflammatory cytokine PIT, pancreatic islet transplantation PVEC, portal vein endothelial cell sICAM, soluble ICAM TNF, tumor necrosis factor TUNEL, TdT-mediated dUTP nick-end labeling assay Pancreatic islet transplantation (PIT) has been validated as treatment for type 1 diabetes ( 1 – 3 ). However...
Journal Articles
Journal: Diabetes
Diabetes 2002;51(5):1529–1534
Published: 01 May 2002
... counted (0.02 mm2). The experiments were done at least in triplicate for each rat. To determine whether renal cell apoptosis was modified by diabetes or rat strain, the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick-end labeling (TUNEL) method for detecting DNA...
Journal Articles
Journal: Diabetes
Diabetes 2002;51(6):1938–1948
Published: 01 June 2002
.... In the hearts of diabetic mice, apoptotic cell death occurred as detected by terminal deoxynucleotidyl transferase–mediated dUTP nick-end labeling (TUNEL) assay. Correspondingly, caspase-3 activation as determined by enzymatic assay and mitochondrial cytochrome c release detected by Western blotting...
Journal Articles
Journal: Diabetes
Diabetes 2003;52(1):165–171
Published: 01 January 2003
... glutathione; STZ, streptozotocin; TUNEL, TdT-mediated dUTP-biotin nick end labeling. ...
Meeting Abstracts
Journal: Diabetes
Diabetes 2000;49(9):1534–1542
Published: 01 September 2000
... the mechanism(s) of CLA reduction of fat mass, female C57BL/6J mice were fed standard semipurified diets (10% fat of total energy) with or without CLA (1% wt/wt). Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick endlabeling (TUNEL) and DNA fragmentation analysis revealed that fat-mass decrease...