Idh2 Deficiency Exacerbates Acrolein-Induced Lung Injury through Mitochondrial Redox Environment Deterioration

Figure 5

The effect of NAC on acrolein-induced toxicity in idh2 knockdown LLC cells. (a) Viability of acrolein-exposed idh2^KD LLC cells in the presence and absence of 1 mM NAC for 2 h was evaluated using MTT assay. Data are presented as the mean ± SD of four independent experiments. versus control cells exposed to acrolein. (b) Immunoblot analysis of apoptosis-associated proteins in idh2^KD LLC cells. β-Actin was used as an internal control. (c) Intracellular ROS levels were measured with DCFH-DA and fluorescence microscopy. (d) Immunoblot analysis of Prx-SO₃ levels in idh2^KD LLC cell lysates. (e) Mitochondrial membrane potential of idh2^KD LLC cells was measured by incorporation of Rh-123 dye into the mitochondria. (f) MitoSox was used to detect mitochondrial ROS generation in idh2^KD LLC cells. MitoSox fluorescence was visualized by a fluorescence microscope. In (c), (e), and (f), the average fluorescence intensity was calculated as previously described [44]. Data are presented as the mean ± SD of four independent experiments. versus WT cells and , versus idh2^KD cells exposed to acrolein.

(a)

(b)

(c)

(d)

(e)

(f)