INSR regulates mitochondrial protein expression and localization. hTCEpi cells and HBECs were treated with siRNA oligonucleotides targeting INSR in KBM for 24 hours. A, To determine the expression of mitochondrial markers in hTCEpi cells, we immunoblotted for VDAC, COX IV, SOD1, and SOD2. Immunoblotting for INSR was used to confirm knockdown. Actin was used as loading control. Immunoblotting showed a decrease in mitochondrial proteins cultured in KBM following INSR knockdown. B, Immunofluorescence was used to analyze expression and localization of the mitochondrial markers COX IV (green) and cytochrome c (Cyto C, red). In hTCEpi cells, COX IV and cytochrome c were decreased in KBM after INSR knockdown. Mitochondrial morphology was also altered, indicating mitochondrial damage. Scale bar: 10 μm. C, Analysis of mitochondrial markers in HBECs did not show a similar effect in KBM. D, Using immunofluorescence, there were no changes in COX IV or cytochrome c expression or localization in HBECs. Scale bar: 10 μm. n = 3