Fig. 2. Rosmarinic acid (RA) induces the nuclear accumulation of nuclear factor erythroid-2-related factor 2 (NRF2), protein kinase B (AKT), and phosphorylation of extracellular signal-regulated kinase (ERK) in human HaCaT keratinocytes. Cells were pretreated with 2.5 μM of RA for 0 to 24 h (3, 6, 12, and 24 h), and the nuclear extract was prepared following the harvest at certain time points. (A) NRF2 activation at different time passages was assessed using western blot analysis using NRF2 and phospho-NRF2 antibodies. TATA-binding protein (TBP) was used as a loading control protein. (B) Intracellular NRF2 localization in cultured cells was assessed via immunocytochemistry using an anti-NRF2 antibody and a FITC-conjugated secondary antibody. Nuclei stained with DAPI appear blue, and NRF2 localization appears green. (C, D) Cells were incubated with 2.5 μM of RA for 0 to 12 h (1, 3, 6, 12 h) intervals, proteins were extracted for western blot, and (C) phospho-AKT, AKT, (D) phospho-ERK, ERK2, were detected using specific antibodies.
© Biomolecules & Therapeutics
