Fig 8.

Treatment of HK-2 with nLDL induces opening of the mitochondrial PTP. HK-2 cells were treated for 24 hrs with 100 μg/ml of nLDL either alone or in combination with 1 μM cyclosporin A (CsA) or 5 μM of the Ca²⁺ ionophore A23187. After that the mtΔΨ, the intramitochondrial Ca²⁺ and the peroxide production were assessed by the specific probes TMRE, Rhod-1 and DCF respectively. In addition cytochrome c was detected by immunofluorescence as detailed under Materials and Methods. To improve visually the data presented, the original fluorescence of Rhod-1 and of the fluorescein isothiocyanate (FITC)-conjugated secondary mAb was digitally rendered in yellow and blue, respectively (by ImageJ 1.38×, NIH, USA, http://rsb.info.nih.gov/ij/) without altering the original pixel intensity scale. A representative LSCM imaging for each probe with a side-by-side statistical evaluation of the fluorescence intensity averaged ± S.E.M. from n= 3 for each condition is shown. Bars inside all the micrographs: 30 μm.