Immunoblot analysis of NHU cell response to TGFβ signalling.

<p>(a) Sub-confluent NHU cells were treated with or without PD153035 (1 µM), TGFβ (2 ng/ml) or SB431542 (3 µM) for 24 h before preparation of cell lysates. Cell extracts (15 µg) were analysed by Western blot analysis as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0051404#s2" target="_blank">Materials and Methods</a>. β-actin was used as an internal loading control. (b) Gel band densities of the Western blots in (E) using the LI-COR Odyssey™ Imaging System and normalised to β-actin to show the changes in phosphorylated protein. The ratio of phosphorylated∶total protein in the control was taken as 1.0, as indicated by the dashed line.