Journal: Antioxidants

Article Title: Acute Oxidative Stress Can Paradoxically Suppress Human NRF2 Protein Synthesis by Inhibiting Global Protein Translation

doi: 10.3390/antiox12091735

Figure Lengend Snippet: H 2 O 2 inhibits global protein synthesis. Cells were treated for 80 min with H 2 O 2 and MnTMPyP as indicated prior to addition of OPP or vehicle. Cells were incubated for an additional 40 min prior to harvest and global protein synthesis analysis. ( A ) Newly synthesized protein signal is AZdye 647 fluorescence. Total protein assessed by Coomassie staining. Images are representative of two biological replicates with two technical replicates each. ( B ) Quantitation of pixel intensity for all replicates, normalized to total protein per lane. * p < 0.001 for H 2 O 2 versus H 2 O 2 + MnTMPyP. No asterisk indicates p > 0.05.

Article Snippet: The reagents in the Click-&-Go Plus 647 OPP Protein Synthesis Assay Kit (Click Chemistry Tools, #1496, Scottsdale, AZ, USA) were stored at 4 °C, except the AZDye 647 Azide Plus, which was dissolved in DMSO and stored in the dark at −20 °C per manufacturer instructions.

Techniques: Incubation, Synthesized, Fluorescence, Staining, Quantitation Assay