p66Shc Mediates Mitochondrial Dysfunction Dependent on PKC Activation in Airway Epithelial Cells Induced by Cigarette Smoke

Figure 5

Pharmacological inhibitors of PKCβ and PKCδ suppressed CSE-induced p66Shc activation and its mitochondrial translocation in airway epithelial cells. The total and phosphorylated protein expression of PKCβ and PKCδ was notably increased in Beas-2b cells exposed to 7.5% CSE for 24 h, and these alterations could be downregulated by 30 min pretreatments with 10 μM LY333531 and 5 μM rottlerin. A representative band is shown for each condition (a), and densitometric analysis of the bands was carried out by Bandscan 5.0 software to quantify the protein expression (b). Further, Western blot analysis revealed that the pretreatments with LY333531 and rottlerin significantly decreased the protein expression of p66Shc and p-p66Shc in the whole cell lysates (c) and suppressed p66Shc mitochondrial translocation and cytochrome c release from the mitochondria (d) in Beas-2b cells incubated with 7.5% CSE for 24 h. All the data are shown as the mean ± SD from three independent experiments. versus the control group and versus the 7.5% CSE-treated group.

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