Figure 3.

Release of nuclear HMGB1 from cells exposed to LTX-315. U2OS cells stably expressing an HMGB-GFP fusion protein were cultured in the indicated conditions with LTX-315, staurosporin (STS), or CCCP. Cells then were fixed and subjected to automatic fluorescence microscopy. Representative images are provided in (a). Quantitative results (means±S.D. of triplicates) are given in (b). Alternatively, parental U2OS cells were subjected to a similar treatment, followed by detection of HMGB1 in the culture supernatant, as shown in (c). Asterisks indicate significant differences (unpaired Student's t-test) with respect to untreated controls. *P<0.05; **P<0.01; ***P<0.001