Journal: Molecules

Article Title: Anti-Inflammatory Effects of Auranamide and Patriscabratine—Mechanisms and In Silico Studies

doi: 10.3390/molecules27154992

Figure Lengend Snippet: The effect of auranamide and patriscabratine on ( A ) NRF2 protein expression; ( B ) HO-1 protein expression; ( C ) NQO1 activity in nuclear extracts of Hepa-1c1c7 cells. The results are expressed as mean ± SD (n = 3). **, p ≤ 0.01; ****, p ≤ 0.0001 compared to LPS Ec + 0.1% DMSO treatment.

Article Snippet: NRF2 ELISA kit (#OKCD09342) was purchased from Aviva Systems Biology, San Diego, CA 92121, USA.

Techniques: Expressing, Activity Assay

Journal: Antioxidants

Article Title: Punicalagin Protects Human Retinal Pigment Epithelium Cells from Ultraviolet Radiation-Induced Oxidative Damage by Activating Nrf2/HO-1 Signaling Pathway and Reducing Apoptosis

doi: 10.3390/antiox9060473

Figure Lengend Snippet: Pre-treatment with 10 µM PUN for 24 h modulates gene expression ( A ), protein synthesis ( B ), and nuclear activation ( C ) of Nrf2 in ARPE-19 cells exposed to UV-A radiation. ( A ) The levels of mRNA expression were measured by RT-PCR. Values are expressed as the mean percentage relative to unirradiated and not pre-treated cells (control 100%) ± SEM of four independent experiment performed in single. ( B ) Nrf2 protein levels were detected by colorimetric cell-based ELISA kit (see “Materials and Methods” section) and were expressed as the mean percentage relative to unirradiated and not pre-treated cells (control 100%) ± SEM of three independent experiment performed in triplicate. ( C ) To quantify Nrf2 activation in the nuclear extract, was utilized a colorimetric assay kit (see “Materials and Methods” section). All OD values were mathematically expressed as a percentage relative to unirradiated and not pre-treated cells (control 100%) ± SEM of three independent experiment performed in duplicate. Differences between mean values (shown in ) were assessed by one-way ANOVA analysis followed by post hoc Newman–Keuls. * = p < 0.05 and *** = p < 0.001 vs. control; °° = p < 0.01 and °°° = p < 0.001 vs. PUN alone; §§ = p < 0.01 and §§§ = p < 0.001 vs. UV-A alone at the same experimental time.

Article Snippet: Intracellular Nrf-2 levels were determined by colorimetric cell-based ELISA kit (LSBio, LifeSpan Biosciences; Seattle, WA, USA).

Techniques: Expressing, Activation Assay, Reverse Transcription Polymerase Chain Reaction, In-Cell ELISA, Colorimetric Assay

Journal: Antioxidants

Article Title: Punicalagin Protects Human Retinal Pigment Epithelium Cells from Ultraviolet Radiation-Induced Oxidative Damage by Activating Nrf2/HO-1 Signaling Pathway and Reducing Apoptosis

doi: 10.3390/antiox9060473

Figure Lengend Snippet: Protective effect of PUN on UV-A-induced apoptosis in ARPE-19 cells. Cells were pre-treated with 10 μM PUN and incubated for 24 h, and then exposed to UV-A radiations for 1, 3, and 5 h. Protein levels of apoptotic hallmarks, Bax ( left-panel ) and Bcl-2 ( right-panel ), were determined by colorimetric cell-based ELISA kits (details in “Materials and Methods” section). Values obtained, normalized to GAPDH, were expressed as a percentage relative to unirradiated and not pre-treated cells (control 100%). Results are from two independent experiments, each including three replicates per experimental group; they are presented as the means ± SEM. One-way ANOVA analysis followed by post hoc Newman–Keuls was carried out. ** = p < 0.01 and *** = p < 0.001 vs. control; °° = p < 0.01 vs. PUN alone; § = p < 0.05 and §§§ = p < 0.001 vs. UV-A alone at the same experimental time.

Article Snippet: Intracellular Nrf-2 levels were determined by colorimetric cell-based ELISA kit (LSBio, LifeSpan Biosciences; Seattle, WA, USA).

Techniques: Incubation, In-Cell ELISA