Journal: Frontiers in Oncology

Article Title: CD47 Enhances Cell Viability and Migration Ability but Inhibits Apoptosis in Endometrial Carcinoma Cells via the PI3K/Akt/mTOR Signaling Pathway

doi: 10.3389/fonc.2020.01525

Figure Lengend Snippet: CD47 up-regulation inhibited apoptosis in HEC-1A and Ishikawa cells. (A,B) Up-regulation of CD47 inhibited cell apoptosis in Ishikawa cell line. (C,D) CD47 overexpression inhibited cell apoptosis in HEC-1A cell line. * p < 0.05.

Article Snippet: The cell cycle of transfected HEC-1A and Ishikawa cells were analyzed using the Cell Cycle and Apoptosis Analysis Kit (Beyotime Biotechnology, Shanghai, China), according to the manufacturer's protocol.

Techniques: Over Expression

Journal: bioRxiv

Article Title: Red yeast rice-derived MKA ameliorates cardiac hypertrophy in hypertensive rats by inhibiting ERK1/2/c-Fos pathway

doi: 10.64898/2026.03.10.710945

Figure Lengend Snippet: Representative images of EDU and TUNEL staining ( A ), EDU positive percent ( B ) and apoptosis percent ( C ) in H9c2 cardiomyocytes treated with different concentrations of MKA (1, 5, 10 μM) and 200 nM Ang II for 24 hours. D-H. Representative Western blots and relative protein expression for β-MHC, p-ERK, t-ERK, c-Fos, and Caspase-3. Data are presented as mean ± SE, comparisons among multiple groups were conducted using one-way analysis of variance (ANOVA) followed by Tukey’s multiple comparison test (n = 4 per group; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001). LM: 1 μM MKA; MM: 5 μM MKA; HM: 10 μM MKA;

Article Snippet: Cells were fixed with 4% paraformaldehyde, permeabilized with 0.3% Triton X-100/PBS (VICMED) for 5 minutes, and processed using the One-step TUNEL In Situ Apoptosis Kit (Red, Elab Fluor® 594) (Elabscience Co., Ltd., Wuhan, E-CK-A322) according to the manufacturer’s instructions.

Techniques: TUNEL Assay, Staining, Western Blot, Expressing, Comparison

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: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Caffeic acid selectively eliminates teratogenic human-induced pluripotent stem cells via apoptotic cell death.

doi: 10.1016/j.phymed.2022.154144

Figure Lengend Snippet: Fig. 3. Increase of caspase activities by CAA treatment in iPSCs. (A) Apoptosis-related proteins were detected by Western blotting in CAA-treated iPSCs and hDFs. Relative band intensities were calculated after normalization to β-actin expression. (B) iPSCs and hDFs were treated with 50 and 100 μM CAA for 24 h, and the caspase-3, −8, and −9 activities were determined. Relative caspase activity compared with CAA-untreated iPSCs and hDFs was expressed as the means ± SD (n = 3). (C) iPSCs were pre-treated with or without 10 μM of caspase-3 inhibitor (z-DEVD), caspase-8 inhibitor (z-IETD), caspase-9 inhibitor (z-LEHD), and pan-caspase inhibitor (z-VAD) for 30 min and then treated with 50 and 100 μM CAA. DMSO was treated as vehicle control. After 24 h, cells were stained with crystal violet solution and relative cell viability compared with CAA-untreated control iPSCs were expressed as the means ± SD (n = 3). **p < 0.01 vs. CAA-untreated control. ##p < 0.01 vs. Inhibitor-untreated control. Scale bar = 100 μM.

Article Snippet: The expression profile of human apoptosis-related proteins in CAA-treated or -untreated iPSCs was analyzed using a Proteome Profiler Human Apoptosis Array Kit A. Kim et al. Phytomedicine 102 (2022) 154144 (ARY009, R&D System, Minneapolis, MN, USA) according to the manufacturer’s instruction.

Techniques: Western Blot, Expressing, Activity Assay, Control, Staining