Figure 2

Extranuclear HMGB1 in CH-induced PH in mice. (A) Translocation of HMGB1 in lung tissue of mice exposed to 24 and 48 h of hypoxia (10% O₂). Release of HMGB1 from the nucleus was assessed by confocal microscopy (red = HMGB1; blue = nuclear stain DAPI (4′,6-diamidino-2-phenylindole); green = autofluorescence to show lung morphology; scale bar = 25 µm; n = 4 mice per group). (B) Representative Western blot against HMGB1 (29 kDa) in serum of C57BL6/J mice exposed to hypoxia (10% O₂ from 2 to 72 h) (n = 4 per time point; serum diluted 1:10; 30 µL loaded per well). (C) Western blot against HMGB1in bronchoalveolar lavage of C57BL6/J mice exposed to hypoxia (10% O₂ from 2 to 72 h) (n = 4 per time point; loading equally 30 µL BAL per well). (D) Expression of HMGB1 in lung tissue of mice exposed to hypoxia (up to 72 h) was assessed by Western blot and normalized to β-actin (43 kDa) (n = 4 per time point).