Fig. 4.

Akt activation in response to control, A549 and SPC-A-1 CM. a Quiescen HUVECs were stimulated with different CM for 15–60 min. Cell lysates were analyzed for Akt activation by Western blot analysis of total and Ser473-phosphorylated Akt (P-Akt) using specific antibodies. Unstimulated cells were parallel control (0 min). b Protein bands were quantified by densitometry, and P-Akt level was calculated for each time point, after normalization to Akt in same sample. Unstimulated basal expression was set as unity. *P < 0.05 and **P < 0.01 versus unstimulated cells (0 min). c Confluent monolayers of HUVECs were pretreated with wortmannin for 20 min, subsequently, the cells were stimulated with A549 CM or treated with vehicle for 30 min. Cell lysates were analyzed for Akt activation by Western blot analysis