Bax expression and co-localization with mitochondria in neutrophils of OSA patients and controls . Bax expression and co-localization with mitochondria was analyzed by confocal laser scanning microscopy in neutrophils of OSA patients and controls in normoxia (Nox) and intermittent hypoxia (IH). The cytoplasmic distribution of mitochondria (red fluorescence) and Bax (green fluorescence) was determined by double immunofluorescence labeling as described in methods. (A, B) Representative images of a control subject (age=38 yr, BMI=25.3 Kg/m2, AHI=6.0 events/hrs, CRP=1.48mg/L, 19% apoptotic neutrophils were detected by Giemsa staining). (A) After 6 hrs of normoxia (Nox) Bax is translocated to and colocalized with mitochondria (yellow and orange). Manders Overlap Coefficient (MOC) >0.6 in neutrophils, (B) IH in-vitro inhibited Bax translocation to the mitochondria. (C) Representative images of a non-obese OSA patient (age=37 yr, BMI=24.7 Kg/m2, AHI=35.0 events/hrs, CRP=1.99 mg/L, 8% apoptotic neutrophils were detected by Giemsa staining) in normoxia and (D) in IH in-vitro. (E) Representative images of an obese OSA patient (age=48 yr, BMI=34.2 Kg/m2, AHI=33.0 events/hrs, CRP=8.09 mg/L, 9% apoptotic neutrophils were detected by Giemsa staining) in normoxia and in (F) IH in-vitro. In obese and non-obese OSA patients Bax was not translocated to the mitochondria; Bax (green) and mitochondria (red) were located separately in normoxia (C, E) and in IH (D, F).