FIGURE 3.

Activation of eicosanoid metabolic program in NPC2-null fibroblasts. Protein expression was detected by Western blotting in the cytoplasmic and nuclear protein fractions isolated from unelicited WT and NPC2-null fibroblasts. The respective proteins were labeled with the specific sets of primary/fluorescently tagged secondary antibodies and were identified on the same membrane by simultaneous dual-color scanning using the high resolution LI-COR Odyssey near-infrared imaging system. The phospho-cPLA2 detection was performed after stripping and re-probing a membrane that was originally stained with the cPLA2 antibodies. Quantitative analysis of COX-2 in nuclear protein fractions from control and NPC2-null cells revealed no statistically significant difference in the relative protein expression levels with the respective mean ± S.E. of 1.22 ± 0.26 (p = 0.4, n = 3).