FIGURE 1.

Itraconazole treatment results in altered glycosylation and increased expression of CD14. RAW 264.7 Mϕs were treated with itraconazole (1 μm) or the respective carrier (DMSO) for 16 h and lysed. Lysates were digested or not with PNGase F (a and b), endo H (b), or sialidase (c) followed by detection of CD14 by Western blotting. Notice that the electrophoretic migration of CD14 after PNGase F or endo H digestion is slightly different, which is consistent with the fact that the first enzyme removes all sugars from the glycoproteins, whereas endo H leaves a GlcNac residue. d, CD14 protein levels were quantitated by densitometry of the Western blot bands and corrected by β-Actin levels. The results represent the average of three separate samples (p < 0.001, t test). e, CD14 mRNA levels were quantitated by real time PCR, corrected by glyceraldehyde-3-phosphate dehydrogenase levels, and normalized to control. The results represent the average of three separate samples (p < 0.001, t test).