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: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Reduction in the developmental potential of intrathymic T cell progenitors with age.

doi: 10.4049/jimmunol.173.1.245

Figure Lengend Snippet: FIGURE 4. Proliferation and survival of ETP from young and old mice. Thymocytes were freshly isolated from young and old mice and stained for ETP in combination with anti-Ki-67 or annexin V labeling to determine their proliferation or survival status, respectively. A, ETP were labeled with anti-Ki-67 or isotype control Abs following fixation and permeabilization. The upper panel shows representative data for single young and old ani- mals. The lower panel indicates data pooled from the indicated number of different aged mice at each time point. B, The level of apoptosis in ETP was evaluated by annexin V labeling. The upper panel shows representative data for single young and old animals. The lower panel indicates data pooled from the indicated number of different aged mice at each time point. Means and SDs are shown for n 2–6 animals as indicated. , p 0.02; , p 0.002; #, p 0.005; ##, p 0.0005.

Article Snippet: The frequency of apoptotic cells in freshly harvested thymocyte populations was assessed by labeling with annexin V using the Apoptosis Detection kit as described by the manufacturer (R&D Systems, Minneapolis, MN) in combination with Abs to various cell surface determinants.

Techniques: Isolation, Staining, Labeling, Control

Journal: Research (Washington, D.C.)

Article Title: Targeting the LSD1-G9a-ER Stress Pathway as a Novel Therapeutic Strategy for Esophageal Squamous Cell Carcinoma.

doi: 10.34133/2022/9814652

Figure Lengend Snippet: Figure 4: Integrative transcriptomics analyses of ESCC cells following pharmacological or genetic inhibition of LSD1 and G9a. (a) RNA-seq analysis was performed on ESCC cells in which either LSD1 expression was knocked down using shRNA or LSD1 was inhibited with SP2509; differentially expressed genes were determined by comparing with ESCC cells expressing a control nonsilencing shRNA (shNC) and vehicle-treated ESCC cells, respectively, and are summarized using Venn diagrams. (b) Gene Set Enrichment Analysis of the overlapping differentially regulated genes in (a). (c) Heatmap of the differentially expressed genes in the “cell death,” “apoptosis,” and “ER stress” pathways. (d–f) Same as (a–c), except G9a was knocked down using shRNA or inhibited with UNC0642. (g–i) Same as (a–c), except both LSD1 and G9a were knocked down with shRNA or both LSD1 and G9a were inhibited with SP2509 and UNC0642.

Article Snippet: The following reagents were also used: hematoxylin and eosin (H&E) staining kit (Beyotime Biotech), TdT in situ apoptosis detection kit (R&D Systems, Minneapolis, MN), TRIzol reagent (Pufei Biotech, Shanghai, China), SYBR Green Supermix (Bimake, Houston, TX), RIPA buffer (Beyotime Biotech), protease inhibitors (Bimake), BCA protein assay kit (Beyotime Biotech), polyvinylidene fluoride (PVDF) membranes (Bio-Rad, Hercules, CA), Pierce ECL System (Thermo Scientific, Waltham, MA), EdU Kit (Ruibo Biotechnology Co., Ltd., Guangzhou, China), cell cycle staining kit (Multi Sciences Biotech, Hangzhou, China), annexin V-FITC/PI apoptosis detection kit (Multi Sciences Biotech), and PrimeScript RT reagent kit (Takara, Kyoto, Japan).

Techniques: Inhibition, RNA Sequencing, Expressing, shRNA, Control

Journal: Research (Washington, D.C.)

Article Title: Targeting the LSD1-G9a-ER Stress Pathway as a Novel Therapeutic Strategy for Esophageal Squamous Cell Carcinoma.

doi: 10.34133/2022/9814652

Figure Lengend Snippet: Figure 5: Inhibiting both LSD1 and G9a in ESCC cells induces S-phase arrest and apoptosis. (a) Summary of the percentage of ESCC cells in the G1, S, or G2 phase after the indicated treatments for 2 days. SP2509 5 μM, UNC0642 5 μM. (b) Western blot analysis of the indicated cell cycle-associated proteins in ESCC cells treated as indicated. (c) EdU staining of ESCC cells treated with vehicle, 3 μM SP2509, 1.2 μM UNC0642, or both for 2 days; the nuclei were counterstained with Hoechst 33342. (d) Apoptosis analysis of ESCC cells after the indicated treatments for 2 days. (e) Western blot analysis of the indicated apoptosis-associated proteins in ESCC cells treated as indicated for 2 days. (f) Quantification of the indicated proteins measured in (e), expressed relative to vehicle-treated cells. (g) Representative images of ESCC cells treated with vehicle, 10 μM SP2509, 10 μM UNC0642, or both for 2 days. (h–k) Representative transmission electron microscopy images of ESCC cells treated as in (g); mitochondria (M) and endoplasmic reticulum (ER) are indicated. The scale bars are 5 μm in (h), 2 μm in (i–k). ∗p < 0:05, ∗∗p < 0:01, and ∗∗∗p < 0:001 (unpaired Student’s t-test).

Article Snippet: The following reagents were also used: hematoxylin and eosin (H&E) staining kit (Beyotime Biotech), TdT in situ apoptosis detection kit (R&D Systems, Minneapolis, MN), TRIzol reagent (Pufei Biotech, Shanghai, China), SYBR Green Supermix (Bimake, Houston, TX), RIPA buffer (Beyotime Biotech), protease inhibitors (Bimake), BCA protein assay kit (Beyotime Biotech), polyvinylidene fluoride (PVDF) membranes (Bio-Rad, Hercules, CA), Pierce ECL System (Thermo Scientific, Waltham, MA), EdU Kit (Ruibo Biotechnology Co., Ltd., Guangzhou, China), cell cycle staining kit (Multi Sciences Biotech, Hangzhou, China), annexin V-FITC/PI apoptosis detection kit (Multi Sciences Biotech), and PrimeScript RT reagent kit (Takara, Kyoto, Japan).

Techniques: Western Blot, Staining, Transmission Assay, Electron Microscopy

Journal: Gene therapy

Article Title: Tumor cells expressing a fusion protein of MULT1 and Fas are rejected in vivo by apoptosis and NK cell activation.

doi: 10.1038/gt.2008.77

Figure Lengend Snippet: Figure 3 Mouse UL16-binding protein-like transcript 1 E (MULT1E)/FasTI induces apoptosis. A total of 1 106 cells of TC-1 and clones L-5, L-7 and L-10 were treated with 1 mg/ml NKG2D/Fc for 16 h. The cells were then analyzed for apoptosis and necrosis using Annexin V assay (a–c) or caspase-3 assay (d) according to the manufacturers’ protocols. (a) An example of the fluorescence-activated cell sorting (FACS) data. (b, c) Summaries of data from three separate experiments. The statistical analyses were conducted between the controls (open bars) and NKG2D/Fc-treated cells (solid bars) using two-way analysis of variance (ANOVA). The difference between NKG2D/Fc-treated L-5 cells and NKG2D/Fc-treated L-10 cells was also compared using Student’s t-test. *Po0.05; **Po0.01 and ***Po0.001.

Article Snippet: Induction of apoptosis in cells expressing the fusion protein To determine if cells expressing the fusion protein can be induced to undergo apoptosis, one million cells of TC-1 and clones L-5, L-7 and L-10 were treated with 1 mg/ml of NKG2D/Fc for 16 h. Apoptosis of the cells was measured using two systems: a TACS Annexin V-FITC Apoptosis Kit (R&D Systems) and a caspase-3 fluorometric assay (R&D Systems).

Techniques: Binding Assay, Clone Assay, Annexin V Assay, Caspase-3 Assay, FACS

Journal: iScience

Article Title: SeSA-HCPT: A dual-targeting agent that induces DNA damage and inhibits repair for castration-resistant prostate cancer therapy

doi: 10.1016/j.isci.2026.114824

Figure Lengend Snippet: SeSA-HCPT induces S-phase cell-cycle arrest and apoptosis in PCa cells Cells were treated with 200 nM SeSA-HCPT, or combined agents (200 nM SeSAHA +400 nM HCPT). DMSO used as control. (A–D) Apoptosis induction in PCa cell lines by the indicated treatments, as determined by flow cytometry. (E–G) Western blot analysis of cleaved caspase-3 levels in PC3, DU145, and LNCaP cells treated with SeSAHA, HCPT, combination therapy, or SeSA-HCPT. (H) Cell cycle distribution analysis of PC3, DU145, and HaCaT cells treated with either SeSA-HCPT alone or the combination therapy. Analysis was performed using flow cytometry. (I–K) Cell-cycle arrest of PC3, DU145, and HaCaT cells by the tested compounds. (L and M) Western blot analysis of cell cycle-related protein expression in PC3 and DU145 cells treated with SeSAHA, HCPT, combination therapy, or SeSA-HCPT. Data are presented as mean ± SD. Statistical significance is denoted as follows: ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: FITC-Annexin V/PI apoptosis kit , Solarbio , Cat#CA1020.

Techniques: Control, Flow Cytometry, Western Blot, Expressing

Journal: Stem Cell Research & Therapy

Article Title: Intravenous injection of BMSCs modulate tsRNA expression and ameliorate lung remodeling in COPD mice

doi: 10.1186/s13287-024-04066-8

Figure Lengend Snippet: Tail vein injection of BMSCs alleviates lung tissue inflammation and apoptosis in COPD model mice. (A) Images of lung parenchyma and peribronchitis H&E staining, along with semi-quantitative analysis of peribronchial inflammation and quantitative analysis of lung parenchymal inflammation. (scale bar = 200 μm; N = 5; * P < 0.05). (B) TUNEL-FITC fluorescence staining images and AI analysis of mouse lung tissue cells. Green fluorescence was FITC-labeled cells with abundant dUTP nick ends, blue fluorescence was DAPI-labeled total cells, and cells labeled with both blue and green fluorescence indicated apoptotic cells. the apoptosis index (AI) wasThe ratio of the number of apoptotic cells to the total number of cells. (scale bar = 100 μm; N = 3; * P < 0.05) Control: naive - Control; COPD: COPD model; VC: COPD model-Tail Vein-Injected BMSCs

Article Snippet: TUNEL staining was conducted to label DNA-damaged apoptotic cells with Annexin V-FITC apoptosis detection kit (Beyotime Cat# C1062M, China).

Techniques: Injection, Staining, TUNEL Assay, Fluorescence, Labeling, Control