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. 2015 Jun 12;290(31):19011–19017. doi: 10.1074/jbc.M115.649210

FIGURE 3.

FIGURE 3.

LPS-induced LKB1 degradation is NO-dependent. A, RAW 264.7 cells were treated with LPS for the indicated time periods. The blot is representative of four blots from four individual experiments. B, RAW 264.7 cells were treated with or without LPS and the iNOS inhibitor SMT (200 μm) for 24 h. Nitrite concentrations in the culture supernatants were measured with the Griess test (n = 6). ***, p < 0.001 versus control. NS, not significant. C, RAW 264.7 cells were treated as described for B, and cell lysates were analyzed for LKB1 expression. The blot is representative of three blots from three individual experiments. D, quantification of LKB1 protein levels as in C (n = 6). ***, p < 0.001 versus control. E, RAW 264.7 cells were transfected with control (si-control) or iNOS (si-iNOS) siRNA (100 nm) for 48 h and treated with LPS for 0, 12, and 24 h. Nitrite concentrations in the culture supernatants were measured with the Griess test (n = 3). *, p < 0.05 versus siRNA control; #, p < 0.01 versus siRNA control. F, RAW 264.7 cells were treated as described for E, and cell lysates were analyzed for LKB1 expression. The blot is representative of three blots from three individual experiments. G, quantification of LKB1 protein levels as in F (n = 3). *, p < 0.05 versus siRNA control. H, RAW 264.7 cells were treated with different doses of GSNO for 24 h. I, quantification of LKB1 protein levels as in H (n = 3). #, p < 0.01 versus control.