Figure 4

S -Nitrosylation of actin by SNAP in PC12 cells. A, B. S-Nitrosylation of actin in PC12 cells by SNAP in the absence (A) and presence (B) of PACAP. PC12 cells (7 × 10⁵ cells/dish) cultured in 6-cm dishes were exposed to 100, 300 or 500 μM SNAP with 10 nM PACAP for 5 min. The cell lysates in HEN buffer were subjected to the biotin-switch assay and analyzed by SDS-PAGE as described in "Methods." C. Quantification of S-nitrosylated actin in PC12 cells. Intensity of bands was quantified by ImageJ, and the extent of S-nitrosylated actin was normalized to total actin. S-Nitrosylated actin by SNAP in the absence (A, open columns) and presence (B, closed columns) of PACAP is expressed as a ratio (mean ± SEM, n = 3) to that at 0 μM SNAP. *P < 0.05 vs.0 μM SNAP without PACAP; ^#P < 0.05 vs.0 μM SNAP with PACAP.