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Fig. 2. Subcellular localization of NOS2 and the various caveolin isoforms in mouse C2C12 myotubes in control experiments (A) and in cells treated with the proinflammatory mixture of LPS/IFN-ß (B). (Left) Myotubes were fixed with paraformaldehyde and methanol, and incubated with antibodies against cav-1, cav-2, cav-3 or NOS2 and either left untreated (A) or treated with LPS/IFN-
for 36 hours (B). (Middle) Subcellular distribution of NOS2 in myotubes untreated (A) or treated with LPS/IFN- for 36 hours (B). The merge signal is depicted on the right. The caveolin fluorescence was visualized by confocal microscopy at an excitation wavelength of 488 nm and is shown in green, whereas the NOS2 was obtained after excitation at 543 nm and is shown in red. Overlap of green and red labeling is depicted in yellow; overlap of green and blue labeling is depicted in light blue; overlap of red and blue is depicted in violet. Scale bar, 50 µM. In all cases, the position of the cell nuclei (blue) was obtained after staining with Hoechst and excitation at 405 nm.
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