Figure 1. NCI-H460 cells express a truncated and catalytically inactive version of Arkadia.
A. Levels of Arkadia and GAPDH mRNA in HaCaT and NCI-H460 cells were assayed by RT-PCR. B. Whole cell extracts from HaCaT and NCI-H460 cells analyzed by Western Blot using an antibody against Arkadia. C. Whole cell extracts from 293T cells transfected with the plasmids indicated were immunoprecipitated (IP) using anti-FLAG beads and Western blotted (WB) using HA and FLAG antibodies. Inputs show expression levels of Arkadia, Smad3, Ski and SnoN. D. NCI-H460 and 293T cells were transfected with the CAGA12-Luciferease reporter and the TK-Renilla control along with plasmids expressing wild-type (WT), C937A, or truncated (1–440) Arkadia. Luciferase activity was normalized to Renilla activity. Means and standard deviations are shown of representative experiments performed in triplicate.