Figure 1

Effects of tunicamycin on ALK expression and phosphorylation and on downstream signaling proteins. Cells were exposed to 500 nM tunicamycin and treated in time course experiments. Expression levels of ALK and phosphorylation of ALK (Tyr1604), AKT (Ser473), ERK1/2 (Thr202/Tyr204), STAT3 (Tyr705) and NPM-ALK (Tyr664) were detected in total cell lysates by Western blot, using corresponding antibodies. α-tubulin was used as sample loading control. Quantification of western blot bands was performed by using ImageJ software [13]. Quantification values for each band were firstly normalized to corresponding values of α-tubulin. Then, normalized values for each time were compared to the corresponding band of the untreated sample (0 hours). Finally the ratio between the phosphorylated form of the proteins and its unphosphorylated counterpart for each experimental time was calculated and reported with standard deviations.