VOLUME 285 (2010) PAGES 30708–30718
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There was an error in the legend to Fig. 5. The legend should read as follows.
Figure 5. A MEK-ERK-RSK pathway is implicated in late signaling events leading to phosphorylation and activation of IKKβ by Ang II in VSMCs. A and B, quiescent VSMCs were pretreated with U0126 (10 μm), PD184352 (2 μm), or DMSO (0.1%) for 30 min before Ang II (100 nm) treatment. Cell extracts were prepared and subjected to immunoblot analysis using the indicated antibodies. One of three independent experiments with similar results is shown. C, quiescent VSMCs were pretreated with U0126 (10 μm) or DMSO (0.1%) for 30 min before Ang II (100 nm) exposure for 15 min. Cell lysates were prepared and analyzed for IKK activity using an in vitro kinase assay. Data shown for DMSO-treated cells were derived from Fig. 2D. One of three independent experiments with similar results is shown. D, densitometric analysis of IKK phosphotransferase activity presented in C. Data are means ± S.E. from three pooled experiments. ##, significantly below the DMSO response (p < 0.01). E, quiescent VSMCs were pretreated with U0126 (10 μm) or DMSO (0.1%) for 30 min before Ang II (100 nm) exposure for 15 min. Nuclear extracts were prepared and subjected to EMSA using NF-κB-specific oligonucleotide as a probe. P50 and P65 represent the use of antibodies to supershift (SS) the inducible DNA-binding complex composed of p65 and p50 subunits. Data shown for DMSO-treated cells were derived from Fig. 2F. One of three independent experiments with similar results is shown. F, densitometric analysis of NF-κB binding activity presented in E. Data are means ± S.E. from three pooled experiments. #, significantly below the DMSO response (p < 0.05). G, quiescent VSMCs were pretreated with BI-D1870 (10 μm) or DMSO (0.1%) for 30 min before Ang II (100 nm) treatment. Cell extracts were prepared and subjected to immunoblot analysis using the indicated antibodies. One of three independent experiments with similar results is shown. H, VSMCs were transfected with a nonsilencing (Ns) RNA duplex or different silencing RNA duplexes that specifically target RSK1, -2, and -3 isoforms. At 48 h post-transfection, cells were serum-starved for 24 h and then exposed to Ang II (100 nm) for the indicated times. Cell extracts were prepared and subjected to immunoblot analysis using the indicated antibodies. One of two independent experiments with similar results is shown. I, quiescent VSMCs were pretreated with Gö6976 (10 μm) and/or with U0126 (10 μm) and/or with DMSO (0.1%) for 30 min before Ang II (100 nm) treatment. Cell extracts were prepared and subjected to immunoblot analysis using the indicated antibodies. One of three independent experiments with similar results is shown.