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. 2014 Apr 28;289(23):16313–16325. doi: 10.1074/jbc.M114.554212

FIGURE 2.

FIGURE 2.

Lysosomal membrane leakage is increased in CHO/CD163 and THP-1 differentiated cells after incubation with Hp 2-2-Hb complex as well as hydrogen peroxide. Lysosomal membrane leakage is presented as the percentage of extralysosomal β-hexosaminidase activity out of the total enzyme activity for each lysosomal preparation. Data shown represent the mean ± S.E. of five independent experiments. A, CHO/CD163 cells. There was an increase in lysosomal injury when the cells were incubated with Hp 2-2-Hb complexes as compared with cells incubated with free Hb (28.7 ± 3.3 versus 8.2 ± 2.1, n = 5, p < 0.001) or Hp 1-1-Hb complexes (28.7 ± 3.3 versus 14.1 ± 3.6, n = 5, p = 0.02). These differences in lysosomal injury were further increased when the cells were preincubated with hydrogen peroxide, and there was significantly higher lysosomal injury associated with Hp 2-2-Hb as compared with Hp 1-1-Hb complexes (39.7 ± 4.6 versus 18.3 ± 2.9, n = 5, p = 0.004). B, there was significantly increased lysosomal leakage in hydrogen peroxide-treated THP-1 cells associated with Hp 2-2-Hb as compared with Hp 1-1-Hb complexes or Hb alone (p < 0.01). * indicates significant differences compared with Hp 1-1 or Hb treatment, and # indicates significant differences between hydrogen peroxides-treated and non-treated conditions.