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. Author manuscript; available in PMC: 2015 Dec 1.
Published in final edited form as: J Cell Physiol. 2014 Dec;229(12):2142–2152. doi: 10.1002/jcp.24677

Figure 4.

Figure 4

Inhibiting PKG negates heparin effects on ERK activity. A7r5 (A, black) and porcine VSMC (A, grey) were synchronized by starvation and stimulated for 30 min with 20 μM PMA. Indicated cells were pretreated for 10 min (A) with heparin at 200 μg/ml or 100 μM 8-Br-cGMP. Indicated cells were incubated with 2 μM Rp-8-pCPT-cGMS for ten min prior to heparin addition. Treated cells were analyzed by Western Blotting for active ERK as described in Methods. Each mean (± SE) represents the activation of ERK2 (n=3) compared to untreated controls (0% activation) and stimulated controls (100% activation). Heparin and 8-Br-cGMP both decreased PMA stimulated ERK activity (p<0.005). The effects of the PKG inhibitor in PMA-activated cells reached significance for the 8-Br-cGMP treated cells (p<0.01). Rp-8-pCPT-cGMS effects on heparin treated cells (**) in these three repeats were not quite at the significance level (p<0.058).