FIGURE 4.
NO does not alter the sGC-α1 Mr distribution profile but causes sGCβ1 to shift its interaction from hsp90 to sGCα1 and back. COS-7 cells that transiently expressed V5-tagged sGC-β1 and Myc-tagged sGCα1 constructs were treated with SNAP (50 μm) or NOC-12 (35 μm), and cell supernatants were generated at indicated times. A and B, Western analysis of gel filtration column fractions showing the sGC-α1 Mr distribution profile in RFL-6 cell supernatants samples. Experimental details are outlined in the legend to Fig. 3, and the same sGC-β1 blots are included here for orientation. C and D, Western analysis of V5-based immunoprecipitations showing bound hsp90, sGC-α1, and sGC-β1 (input 20%) retained on the beads. E, band intensities versus time of NO treatment for the hsp90 and sGC-α1 bound to sGC-β1 as determined from D. Data are representative of three replica experiments.