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Fig. 4. Treatment of myotubes with ·NO donors induce changes in the levels of cav-3, but not of cav-1 or cav-2. C2C12 myoblasts were differentiated into myotubes and incubated with LPS (2 µg ml-1) plus IFN-
(100 U ml-1), the ·NO donor DETA-NONOate (100 µM), the ·NO inhibitors 1400W (100 µM) and nitro-Arg (100 µM) for 36 h (cav-1 and cav-2) or 48 h (cav-3) in different combinations. The cells were scraped and the changes in cav-1 (white bars), cav-2 (single-slashed bars) and cav-3 (double-slashed bars) protein levels were determined by immunodetection (A). In addition, activated C2C12 myotubes were incubated with the ·NO donor DETA NONOate, the lipophilic cGMP analog 8-bromo-cGMP (20 µM), with the soluble guanylate cyclase inhibitor ODQ (10 µM) or with cycloheximide (CHX; 10 µM) in various combinations for 48 hours, and the levels of cav-3 were determined by immunodetection (B). In every case, the total amount of protein loaded was similar, as judged by ß-tubulin quantification. *P<0.001 vs the corresponding condition in the absence of ·NO donor. Averaged results are shown, being representative of at least three independent experiments (±s.d.).
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