Research Article

4-Hydroxy-2-nonenal Alkylated and Peroxynitrite Nitrated Proteins Localize to the Fused Mitochondria in Malpighian Epithelial Cells of Type IV Collagen Drosophila Mutants

Figure 3

Cytoplasmic membrane-associated HNE-modified proteins in mutants. Color code: protein-HNE adducts, green; cytoplasmic membrane red; nuclei, blue. (a) Protein-HNE adducts in wild-type flies incubated at 20°C. (b) Overlay with membrane staining. (c) Protein-HNE adducts in wild-type flies incubated at 29°C. (d) Merged with membrane staining. ((e), (f), (g), (h)) Representative mutant, incubated at 20 or 29°C presented in the same order as in upper row. White arrows in (b), (d), (f), and (h). Regions presented in higher magnification in (i), (j), (k), and (l), respectively. White arrows in (i), (j), (k), and (l) show association of the cytoplasmic membrane with alkylated proteins. Note the notorious infiltration of HNE-modified proteins into the membrane in (l), which occurs at a less extent in (k). The membrane of wild-type animals is free of alkylated proteins (i), while they associate closely with the membrane in the mutant (j), incubated at permissive temperature.