Fig. 2

Inhibitory effect of LL28 on the activation of both IGF1R and Src. a A549 cells were treated with linsitinib (1 μM), dasatinib (100 nM), or LL28 (1 μM) for 4 h. Before harvesting, cells were stimulated with FBS for 20 min. The expression of total and phosphorylated IGF1R and Src was evaluated by Western blot analysis. b and c A549, H1299, and H460 cells were treated with LL28 (0.1 and 1 μM) for 8 h (b and c) or 1.5 days (c). b The expression of total and phosphorylated IGF1R and Src was evaluated by Western blot analysis. c The expression of the total and phosphorylated forms of several kinases was evaluated by Western blot analysis. d Total cell lysates of A549 cells treated with LL28 for 8 h were immunoprecipitated with anti-IGF1R or anti-IR antibodies. The immunoprecipitants were further subjected to Western blot analysis using anti-pTyr, anti-IGF1R, and anti-IR antibodies. e R- cells were treated with LL28 (0.1 and 1 μM) for 8 h. The expression of total and phosphorylated IGF1R and Src was determined by Western blot analysis. f A549 cells were treated with linsitinib (1 μM) or dasatinib (100 nM) for 1 day. The expression of total and phosphorylated IGF1R and Src was evaluated by Western blot analysis. g A549, H1299, and H460 cells were treated with LL28 (0.1 μM) for 1, 3, and 5 days. The expression of total and phosphorylated IGF1R and Src was evaluated by Western blot analysis. Con: control; Lin: linsitinib; Das: dasatinib