Fig. 4.

PTM increases CRT exposure and cytoplasmic translocation of HMGB1 from the nucleus of LLCs. (a) Surface-exposed CRT (green) and (b) translocated HMGB1 (green) in LLCs treated with medium (CTR), 25% PTM, pretreated 10 mM NAC, or 0.1 mM 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide(cPTIO) and subsequently with 25% PTM for 12 h. Confocal images indicate stained nuclei (blue), and CRT or HMGB1 (green); scale bar, 10 μm. Arrows indicate increased fluorescence intensity of HMGB1 in the nuclei. Quantitative co-localization of HMGB1 expressed as nuclear (Hoechst) (n) per cell. HMGB1 staining intensity was analyzed using Zen blue lite. Data are represented by the mean ± standard error of the difference between two means (SED) and calculated using a two-tailed unpaired Student’s t-test. This experiment was repeated thrice; ***p < 0.001, and ****p < 0.0001.n.s., not significant