Figure 4

Downregulation of the IGF-1/insulin pathways by klotho in lung cancer cell line A549. A549 cells were transfected with either MYC-KL or control vector pCMV6. After 24 hr, cells were serum-starved for 24 hr and treated with IGF-1 (10 min, 25 nM) or insulin (10 min, 100 nM). Following treatment, cells were harvested and proteins were resolved and immunoblotted using antibodies either directed against phospho (P) and total (T) IGF-1R or phospho (P) and total (T) insulin-R (IR). Similar treatment was done when silenced the klotho of the cells using sh-2 or control shRNAc. Data shown are the mean results ± SD of a representative experiment performed in triplicate (n = 3), *indicates p < 0.01.