Journal: PLoS ONE

Article Title: Potent Anti-Cancer Effect of 3′-Hydroxypterostilbene in Human Colon Xenograft Tumors

doi: 10.1371/journal.pone.0111814

Figure Lengend Snippet: Cells were treated with various concentrations (5, 10, 25 and 50 µM) of pterostilbene or 3′-hydroxypterostilbene for 24 h. (A) Determination of sub-G1 cells in COLO 205 cells by flow cytometry after PI staining as described in the . (B, C) After treatment, total cell lysates were prepared from COLO 205 cells and the cleavage of PARP, DFF-45, pro-caspase 8 and pro-caspase 9 were analyzed by Western blotting. (D) Kinetics of caspase activation in COLO 205 cells. Cells were treated with 25 and 50 µM of pterostilbene or 3′-hydroxypterostilbene for 24 h. Caspase activities were analyzed as described in the . (E) Cells were treated with 50 µM of pterostilbene or 3′-hydroxypterostilbene for 15 min. Mitochondrial membrane potential and ROS production were stained with DiOC6 (40 nM) and DCFH-DA (20 µM) and measured by flow cytometry. The values are expressed as means ±SE of triplicate tests. * P <0.05 indicates statistically significant difference from the pterostilbene-treated group.

Article Snippet: The PARP, DFF-45, LC-3 I/II, p-mTOR (Ser 2448 ), mTOR, p-P70S6K (Thr 398 ), p-P70S6K (Ser 371 ), p-Akt (Ser473), Akt, PI3K, p-ERK1/2 (Thr202/Tyr204), p-JNK1/2 (Thr183/Tyr185), JNK1/2, p-p38 (Thr180/Tyr182), p38 and MMP-9 antibodies were purchased from Cell Signaling Technology (Beverly, MA).

Techniques: Flow Cytometry, Staining, Western Blot, Activation Assay

Journal: International Journal of Molecular Sciences

Article Title: Antileukemic Natural Product Induced Both Apoptotic and Pyroptotic Programmed Cell Death and Differentiation Effect

doi: 10.3390/ijms222011239

Figure Lengend Snippet: Determination of ardisianone-induced cell death in HL-60 cells. ( A ) Cells were treated with or without ardisianone at the indicated concentration and time. After the treatment, the cells were stained with Annexin V-PI to analyze apoptosis using flow cytometric analysis. ( B ) Cells were treated with graded concentrations of ardisianone for 24 h. After the treatment, the cells were fixed and stained with propidium iodide to analyze DNA content by FACScan flow cytometer, and quantitative analysis of sub-G1 (apoptosis) population was performed. ( C ) Cells were treated with or without ardisianone (2 μM) for 3 to 24 h. Total protein was extracted and subjected to SDS–PAGE for the detection of apoptotic DFF45/ICAD. Protein expression was quantified using Bio-Rad Image Lab TM Software. * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with respective controls.

Article Snippet: Antibodies of DFF45/DFF35, TNFR1, TNFR2, Fas, DR5, Bid, cleaved caspase-1, caspase-3, caspase-5, caspase-8, gasdermin D, cIAP1, cIAP2, survivin, HMGB1, and γH2A.X Ser139 were obtained from Cell Signaling Technologies (Boston, MA, USA).

Techniques: Concentration Assay, Staining, Flow Cytometry, SDS Page, Expressing, Software

Journal: PLoS ONE

Article Title: Muscadine Grape Skin Extract Induces an Unfolded Protein Response-Mediated Autophagy in Prostate Cancer Cells: A TMT-Based Quantitative Proteomic Analysis

doi: 10.1371/journal.pone.0164115

Figure Lengend Snippet: Western blot analysis in C4-2 cells treated with 20 μg/ml MSKE as compared to ethanol-treated controls. As a loading control total protein from Ponceau S staining was assessed. Expression of ER stress markers IRE1-alpha and GRP78 and pro-apoptotic markers DFF45, PARP and caspase-12 was analyzed and quantification of western blot analysis performed using Image J, NIH. The standard deviation was used to assess data dispersion.

Article Snippet: The following primary antibodies were used at a 1:1000 dilution LC3B, cleaved caspase-3, cleaved caspase-7, BAX, and BCL2 (Cell Signaling Technologies Danvers, MA.), anti-caspase-12, Ire-1α, GRP78, DFF45, and PARP (Abcam, Cambridge, MA).

Techniques: Western Blot, Staining, Expressing, Standard Deviation