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. Author manuscript; available in PMC: 2014 Feb 3.
Published in final edited form as: J Cereb Blood Flow Metab. 2009 Mar 25;29(6):1084–1098. doi: 10.1038/jcbfm.2009.32

Figure 5.

Figure 5

Expression of Smad proteins in brain microvessels during CIP in the presence and absence of an ALK5 inhibitor. Western blot analysis of microvessels isolated from rats treated with saline or λ-carrageenan in the presence and absence of SB431542, a selective ALK5 inhibitor. Cytoplasmic and nuclear extracts from rat brain microvessels (20 µg) were resolved on a 10% sodium dodecyl sulfate–polyacrylamide gel and transferred to a PVDF membrane. Samples were analyzed for expression of p-Smad2 and p-Smad3 (A) as well as p-Smad1/5/8 (B) and Smad4 (C). P-Smad2 was detected using the polyclonal antibody Ser465/467 (1:500 dilution), p-Smad3 was detected using the polyclonal antibody Ser423/425 (1:500 dilution), pSmad1/5/8 was detected using the polyclonal antibody Ser463/465 (1:500 dilution), and Smad4 was detected using the polyclonal anti-Smad4 antibody (1:500 dilution). Relative levels of Smad proteins were determined by densitometric analysis and the nucleus/cytoplasm ratio was calculated. Results are expressed as mean ± s.d. of three separate experiments. Asterisks represent data points that are significantly different from control.