Journal: Current Issues in Molecular Biology

Article Title: Effect of Soy Isoflavone on Prostate Cancer Cell Apoptosis Through Inhibition of STAT3, ERK, and AKT

doi: 10.3390/cimb46110743

Figure Lengend Snippet: Caspase-3 activation by genistein in HPrEC and DU145 cells. ( A – C ) Cells were treated with increasing concentrations (0–100 μM) of genistein for 48 h. Caspase-3/7 activity was determined using an ApoTox-Glo TM Triplex Assay ( A ). Cell lysates were subsequently analyzed by Western blotting with antibodies against procaspase-3, cleaved caspase-3, PARP, Bax, Bcl-2, and β-actin ( B ). The percentage of apoptotic cells after annexin V-PE binding was quantified by a Muse cell analyzer ( C ). ( D – F ) Cells underwent treatment with genistein (100 μM) for 48 h in the presence or absence of Z-VAD (10 μM). Caspase-3/7 activity was determined using the ApoTox-Glo TM Triplex Assay ( D ). Cell lysates were then analyzed with Western blotting for procaspase-3, cleaved caspase-3, PARP, and β-actin ( E ). The percentage of apoptotic cells after annexin V-PE binding was determined using a Muse cell analyzer ( F ). Error bars represent mean ± S.D. from three independent experiments. * p < 0.05 was compared to respective controls. # p < 0.05 was compared to Z-VAD (−) groups. GEN: genistein, HPrEC: human prostate epithelial cell line, and Conc.: concentration.

Article Snippet: Cells were harvested, resuspended in the culture medium, mixed with Muse TM Annexin V & Dead Cell reagent, and analysis was performed using a Muse TM Cell Analyzer (Merck Milipore, Billerica, MA, USA).

Techniques: Activation Assay, Activity Assay, Western Blot, Binding Assay, Concentration Assay

Journal: Pharmaceuticals

Article Title: Novel Therapeutic Effects of Euphorbia heterophylla L. Methanol Extracts in Macular Degeneration Caused by Blue Light in A2E-Laden ARPE-19 Cells and Retina of BALB/c Mice

doi: 10.3390/ph17091193

Figure Lengend Snippet: Level of cell deaths parameters in the MEE + A2E + BL-treated ARPE-19 cells. ( A ) Relative level of cell viability and ( B ) morphology of the MEE + A2E + BL-treated ARPE-19 cells. After pretreatment with three different dosages of the MEE for 24 h, their viability was analyzed using the MTT assay. The MTT assays were performed on samples from two to three wells per group, and the optical density was measured twice for each well. ( C ) The number of specific cells stained with Annexin V and 7-AAD. After staining, the number of stained cells was analyzed using a Muse Cell Analyzer. Annexin V and 7-AAD staining were performed from two to three wells per group, and the number of specific cells was counted twice for each well. The red lines represented four different stages of cell death, while individual cell was marked with a red dot. ( D ) Protein expression level of Bax, Bcl-2, Cas-3, and Cleaved Cas-3. The expression level of each protein was normalized to β-actin. The cell lysates were prepared from two to three dishes per group and the Western blots were analyzed twice for each sample. Data are reported as the mean ± SD. * p < 0.05 vs. No group. # p < 0.05 vs. Vehicle + A2E + BL-treated group. Abbreviations: MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; 7-AAD, 7-aminoactinomycin D; Bax, Bcl-2-associated X protein; Bcl-2, B-cell lymphoma 2; Cas-3, Caspase-3; MEE, methanol extracts of Euphorbia heterophylla L.; A2E, N-retinylidene-N-retinylethanolamine; BL, blue light; ARPE, arising retinal pigment epithelia.

Article Snippet: These cells were analyzed using the Muse TM Cell Analyzer (Millipore Co.), and their distribution was presented as the population of live and apoptotic cells.

Techniques: MTT Assay, Staining, Expressing, Western Blot