Journal:

Article Title: Skin-stage Schistosomula of Schistosoma mansoni Produce an Apoptosis-inducing Factor That Can Cause Apoptosis of T Cells *

doi: 10.1074/jbc.M201344200

Figure Lengend Snippet: Following incubation, apoptosis was measured by annexin V and propidium iodide staining and counting 500 –1000 cells under a fluorescence microscope (A), by measuring caspase-3 activity in various subset of lymphocytes using PhiPhiLux substrate and counting 500 –1000 cells under a fluorescence microscope (B), and by evaluating DNA fragmentation in different subsets after treatment with DNA digestion buffer and separation on a 1% agarose gel (C). Data presented are representative of one of three to five similar experiments using five to seven mice per group in each experiment. A and B show means ± S.D. of the percentage of positive cells. * and **, p < 0.05 and p < 0.01 compared with the medium control, respectively.

Article Snippet: Apoptotic cells around the parasites in the skin of mice were evaluated by TUNEL staining using an in situ apoptosis detection kit (R&D Systems, Minneapolis, MN) following the instructions of the manufacturer.

Techniques: Incubation, Staining, Fluorescence, Microscopy, Activity Assay, Agarose Gel Electrophoresis, Control

Journal:

Article Title: Skin-stage Schistosomula of Schistosoma mansoni Produce an Apoptosis-inducing Factor That Can Cause Apoptosis of T Cells *

doi: 10.1074/jbc.M201344200

Figure Lengend Snippet: 1 × 106 Thy1.2+ cells isolated from skin-draining lymph nodes of naive mice were cultured with 60 μg/ml ES products or ES products plus 5 μg/ml recombinant Fas-Fc fusion protein for 24 h. Apoptosis was evaluated by determining the percentage of annexin V-positive cells. *, p < 0.01 compared with the medium control.

Article Snippet: Apoptotic cells around the parasites in the skin of mice were evaluated by TUNEL staining using an in situ apoptosis detection kit (R&D Systems, Minneapolis, MN) following the instructions of the manufacturer.

Techniques: Isolation, Cell Culture, Recombinant, Control

Journal:

Article Title: Skin-stage Schistosomula of Schistosoma mansoni Produce an Apoptosis-inducing Factor That Can Cause Apoptosis of T Cells *

doi: 10.1074/jbc.M201344200

Figure Lengend Snippet: ES products were divided into three fractions, fraction 1 (Fr 1; <3 kDa), fraction 2 (Fr 2; 3–30 kDa), and fraction 3 (Fr 3; >30 kDa). Bands in fraction 2 were again separated by gel filtration. Each peak (at 60 μg/ml) was then tested for its ability to induce apoptosis of T cells (isolated from the skin-draining lymph nodes of naive mice) by staining for annexin V. Only two peaks (23 and 28 kDa) are shown. About 500 cells were counted under a fluorescence microscope. Values are representative of one of three similar experiments and are means ± S.D. of the percentage of positive cells. * and **, p < 0.05 and p < 0.001 compared with the medium control (M), respectively. ES products from normal schistosomula were used as a positive control.

Article Snippet: Apoptotic cells around the parasites in the skin of mice were evaluated by TUNEL staining using an in situ apoptosis detection kit (R&D Systems, Minneapolis, MN) following the instructions of the manufacturer.

Techniques: Filtration, Isolation, Staining, Fluorescence, Microscopy, Control, Positive Control

Journal: Biochimica et biophysica acta. Molecular basis of disease

Article Title: p85α deficiency alleviates ischemia-reperfusion injury by promoting cardiomyocyte survival.

doi: 10.1016/j.bbadis.2024.167318

Figure Lengend Snippet: Fig. 7. The ubiquitin proteasome pathway, apoptosis, mitochondrial function, and inflammation are enriched in the transcriptomic analysis of human failing myocardium. (A) GO terms enrichment analysis based on differentially-expressed genes (DEGs). DEGs were identified using a significance threshold of P.adj < 0.05 in the Zenodo DataSets (ID: 4114617), comparing HFrEF versus control. Additionally, DEGs were identified using the same threshold in the GEO DataSets (ID: GSE116250), comparing ICM versus NF. HFrEF, heart failure with reduced ejection fraction; ICM, ischemic cardiomyopathy; NF, non-failing donors. (B) KEGG pathway enrichment analysis of DEGs identified in (A). (C) Schematic representation of how p85α signaling aggravates myocardial ischemia-reperfusion injury. Left: in WT mice, p85α protein is increased after myocardial I/R injury. The elevation of p85α results in reduced phosphorylation levels of Akt and GSK3β, as well as decreased abundance of Bcl-x(L). Consequently, apoptosis is aggravated and inflammation genes are activated. In addition, p85α leads to mitochondrial dysfunction, ultimately exacerbating myocardial I/R injury. Right: in p85α−/−mice, the lack of p85α increases Akt and GSK3β phosphorylation along with upregulation of Bcl-x(L) expression, subsequently improving cardiac inflammation and apoptosis. Moreover, p85α deficiency promotes mitochondrial function and increases mtDNA content through the upregulation of ERRα, and PGC-1α expression. The E3 ligase MG53 mediates the ubiquitin-dependent degradation of p85α protein, highlighting the importance of posttranslational modification in regulating p85α expression. Together, knocking out p85α or promoting p85α ubiquitin-dependent degradation mediated by MG53 has protective effects against myocardial I/R injury.

Article Snippet: To evaluate DNA fragmentation in myocardial sections, we used the CardioTACSTM in situ apoptosis detection kit (#TA5353; R&D Systems Inc., Minneapolis, MN), following previously described methods [11,12].

Techniques: Ubiquitin Proteomics, Control, Phospho-proteomics, Expressing, Modification

Journal: Redox Biology

Article Title: Redox homeostasis and cell cycle activation mediate beta-cell mass expansion in aged, diabetes-prone mice under metabolic stress conditions: Role of thioredoxin-interacting protein (TXNIP)

doi: 10.1016/j.redox.2020.101748

Figure Lengend Snippet: Morphology and TUNEL-assay of young and aged NZO mice before and after carbohydrate feeding. (A) Representative images (20x magnification) showing multi-color immunofluorescent staining in longitudinal, serial sections from young and aged NZO mice at indicated time points. Green = insulin, red = glucagon, yellow = somatostatin, blue = DAPI. Scale bars, 50 μm. (B) Beta-cell mass, (C) total islet number per slide and (D) total nuclei number per islet in young (green) and aged (grey) NZOs before and 2 as well as 21 days after + CH feeding (n = 8–10). (E) Quantification of TUNEL + -nuclei, given as percentage of total nuclei number within the insulin + -area (n = 4). Results are represented as mean values ± SD. Statistical significance was assessed by two-way ANOVA with Sidak's multiple comparison test, a,b,c,e p < 0.05 (a = significant to young NZO 0, b = significant to aged NZO 0, c = significant to young NZO 2, e = significant to young NZO 21). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Apoptotic cells in pancreatic tissue sections were detected via TUNEL assay - TdT In Situ Apoptosis Detection Kit Fluorescein (R&D Systems, Minneapolis; USA), according to supplier's instructions.

Techniques: TUNEL Assay, Staining, Comparison