Effect of PDGF-BB (20 ng/ml) on ERK1/2 and NO signaling.

<p>A) Time course of ERK1/2 activation in response to PDGF, B&C) Time course of generation of intracellular nitric oxide in response to PDGF, D) Activation of eNOS through phosphorylation of Ser¹¹⁷⁷ residue in response to PDGF and E) PDGF-induced expression of iNOS. Bar graphs summarize data for a minimum of four independent experiments. *P<0.05, **P<0.01 and ***P<0.001 versus control.</p

Role of ERK1/2-NO axis on PDGF-induced proliferation of RASMCs and effect of <i>G lutea</i> on PDGF induced NO index.

<p>A) Effect of ERK inhibitor 328000 (1 µmole/L) on PDGF induced cell proliferation, B) Effect of NOS inhibitor L-NAME (10 µmole/L) on PDGF induced cell proliferation, C) Effect of <i>G lutea</i> extract on generation of cellular NO in response to PDGF. *P<0.05 versus control and †<0.05, ††<0.01 and †††<0.001 versus corresponding PDGF treatment.</p

Effect of isovitexin on PDGF- induced RASMC proliferation.

<p>A) Docking analysis revealing binding pocket of isovitexin on MEK1, B) Effect of isovitexin (0.5–10 µmol/L) on PDGF induced ERK1/2 activation and cell cycle progress and C) Effect of isovitexin on PDGF induced cell proliferation measured through alamar blue assay. Bar graphs summarize data for a minimum of three independent experiments. **P<0.01 versus control and ††<0.01 and †††<0.001 versus PDGF treatment.</p

Effect of <i>G lutea</i> extract on PDGF-induced cell signaling in RASMCs.

<p>A) PDGFR-β phosphorylation, B) ERK1/2 activation, C) Phosphorylation of IKKα and D&E) Representative blot and bar graph indicating expression of cyclin D1, PCNA and iNOS for a minimum of three independent experiments. *<0.05, **P<0.01 and ***P<0.001 versus control and ††<0.01 and †††<0.001 versus PDGF treatment.</p

Predicted binding energies for <i>G lutea</i> constituents and known MEK1. inhibitors.

<p>Predicted binding energies for <i>G lutea</i> constituents and known MEK1. inhibitors.</p

IC₅₀ values for the <i>G. lutea</i> extract (1 mg/ml) on smooth muscle cells.

<p>A) Primary cultures of rat aortic smooth muscle cells (RASMCs), B) Rat aortic smooth muscle cell line A7r5 and C) Human aortic smooth muscle cell line ATCC-CRL-1999.</p

Cell cycle analysis of RASMCs.

<p>A) Representative flow cytogram depicting cells in various stages of cell cycle upon treatment with PDGF-BB (20 ng/ml) in presence and absence of <i>G. lutea</i> extract, and B) Bar graph summarizing data for four independent experiments. ***P<0.001 versus control, ††<0.01 and †††<0.001 versus PDGF treatment.</p

Summary figure depicting the pathway blocked by <i>G lutea</i> extract to prevent PDGF-induced RASMC proliferation.

<p>Summary figure depicting the pathway blocked by <i>G lutea</i> extract to prevent PDGF-induced RASMC proliferation.</p

Effect of <i>G. lutea</i> extract (1 mg/ml) on proliferation and apoptosis.

<p>A) Effect of extract on PDGF-BB (20 ng/ml, 24 hours) induced proliferation of primary cultures of rat aortic smooth muscle cells (RASMCs), B) A7r5 and C) percentage of apoptotic RASMCs in response to experimental treatments. *P<0.05 and ***P<0.001 versus control and †P<0.05 and †††<0.001 versus PDGF treatment.</p