Figure 2: NPM1 translocation is elicited by nucleolar oxidation.
From: A redox mechanism underlying nucleolar stress sensing by nucleophosmin
(a) Endogenous NPM1 translocation under various cellular stresses±NAC pretreatment (as in Fig. 1b). Representative images (left) and the RFI of the nucleolar NPM1 (right) were displayed. n=55 cells. Unpaired t-test, ***P<0.001 with respect to treated versus untreated or −NAC versus +NAC cells. The nucleolar and nucleoplasmic line profiles of NPM1 were analysed in Supplementary Fig. 2a. (b) EGFP-NPM1 translocation upon H2O2 (500 μM) exposure, visualized by live-cell imaging. Representative images at indicated times were displayed. (c,d) RFI changes of nucleolar EGFP-NPM1 per minute upon three doses of H2O2 treatments or H2O2 (500 μM) plus pretreatment of NAC. (e) EGFP-NPM1 translocation upon H2O2 (500 μM) exposure followed by the addition of 5 mM DTT after 20 min. (f) EGFP-NPM1 translocation and nucleolar redox changes upon H2O2 (500 μM) exposure±DTT pretreatment in the same cells. Concurrent changes of relative 405/488 nm ratio values reflecting redox state (left y axis) and RFI of nucleolar EGFP-NPM1 (right y axis) per minute were plotted. Scale bars, 10 μm (a) and 5 μm (b,e,f). Data were represented as mean±s.e.m.
