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