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. 2019 May 20;14(5):e0216668. doi: 10.1371/journal.pone.0216668

Fig 4. DEX activates a delayed response in WT cells but not in AT cells.

Fig 4

Fyn nuclear import and NRF2 phosphorylation were detected in WT and AT cells. A) Western blot analysis for Fyn in the cytosolic/nuclear fractions of WT (238RM) and AT (AT28RM, AT50RM, AT129RM) cells treated as reported in Fig 2. HPRT1 and LAMIN A served as a loading control for the cytosolic/nuclear extracts, respectively. B) Quantification of the relative amount of Fyn in the cytosol and nuclei of WT and AT cells is shown. C) Western blot analysis for phosphorylated NRF2 and total NRF2 in the WT (238RM) and AT (AT129RM) cell lines, treated or not with DEX as above. After nuclei separation and lysis, NRF2 was immunoprecipitated and probed with either anti phospho-tyrosine or anti-NRF2. LAMIN A served as a loading control; HPRT1 was used to assess the lack of cross contamination and the consequent nuclear localization of NRF2. D) Quantification of the p-NRF2/NRF2 ratio. Blots shown are representative and the histograms are the means and SEM of four independent experiments (Wilcoxon signed rand test; *two-tailed p-values<0.05).