tacs 2 tdt dab in situ apoptosis detection kit  (Bio-Techne corporation)

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: Oxidative Medicine and Cellular Longevity

Article Title: Geraniin Protects against Cerebral Ischemia/Reperfusion Injury by Suppressing Oxidative Stress and Neuronal Apoptosis via Regulation of the Nrf2/HO-1 Pathway

doi: 10.1155/2022/2152746

Figure Lengend Snippet: The effect of geraniin on neuronal apoptosis 72 h after MCAO in rats. (a–c) Effect of geraniin on MCAO/R-induced neuronal apoptosis in cerebral cortex and hippocampal sections, as determined by TUNEL staining (40x, scale bar = 20 μ m) ( n = 4). The white arrows indicate TUNEL-positive cells. (d–h) The expression levels of Bax, Bcl-2, cleaved caspase-3, and caspase-3 were measured by Western blot analysis ( n = 6). The data are expressed as the means ± SEM. ▲▲ P < 0.01 vs. the sham group; ∗ P < 0.05, ∗∗ P < 0.01 vs. the MCAO/R group.

Article Snippet: A TdT In Situ Apoptosis Detection Kit was purchased from R&D Systems, Inc. (Minneapolis, USA).

Techniques: TUNEL Assay, Staining, Expressing, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Geraniin Protects against Cerebral Ischemia/Reperfusion Injury by Suppressing Oxidative Stress and Neuronal Apoptosis via Regulation of the Nrf2/HO-1 Pathway

doi: 10.1155/2022/2152746

Figure Lengend Snippet: The effect of geraniin on OGD/R-induced apoptosis of PC12 cells. (a, b) Effect of geraniin on the OGD/R-induced cell apoptosis rate, as determined by flow cytometry. (c–f) The protein expression of Bax, Bcl-2, caspase-3, and cleaved caspase-3 was measured by Western blot analysis. The data are expressed as the means ± SEM ( n = 8). ▲▲ P < 0.01 vs. the control group; ∗ P < 0.05, ∗∗ P < 0.01 vs. the OGD/R group.

Article Snippet: A TdT In Situ Apoptosis Detection Kit was purchased from R&D Systems, Inc. (Minneapolis, USA).

Techniques: Flow Cytometry, Expressing, Western Blot, Control