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. Author manuscript; available in PMC: 2015 Jan 1.
Published in final edited form as: Cell Biochem Biophys. 2014 Jan;68(1):10.1007/s12013-013-9677-6. doi: 10.1007/s12013-013-9677-6

Figure 1. Phosphorylation of M-RIP and increased association of M-RIP with MYPT1 by activators of PKG.

Figure 1

(A) Gastric smooth muscle cells labeled with 32P were incubated with GSNO (10 μM) in the presence or absence of PKG inhibitor Rp-cGMPS (RpG, 10 μM), or 8-cCPT-cGMP (8pCPT, 10 μM) for 10 min. Immunoprecipitates derived from 500 μg of protein using M-RIP antibody were separated on SDS-PAGE. Bands corresponding to [32P]M-RIP were identified using autoradiography. Radioactivity in the bands is expressed as cpm. Immunoblot analysis showed equal amounts of loaded protein. Rp-cGMPS (10 μM) alone had no effect on basal M-RIP phosphorylation (483±67 cpm/mg protein). Values are means ± SE of 4 experiments. **P < 0.01, significant increase of M-RIP phosphorylation. (B) Dispersed smooth muscle cells were incubated with GSNO (10 μM) in the presence or absence of PKG inhibitor Rp-cGMPS (RpG (10 μM), or 8-cCPT-cGMP (8pCPT, 10 μM) for 10 min. Immunoprecipitates derived from 500 μg of protein using MYPT1 antibody were separated on SDS-PAGE and immunoblotted using M-RIP antibody. Rp-cGMPS (10 μM) alone had no effect on basal association of M-RIP with MYPT1(data not shown). Values are means ± SE of 4 experiments. **P < 0.01, significant increase in M-RIP and MYPT1 association.