LPS-induced degradation of DOK3 mediates ERK activation.

<p>(A) RAW264.7 macrophages, transduced with 23 nt plus hairpin shRNA designed to knock down DOK3 or a control luciferase knock down, were lysed and decreased DOK3 expression was confirmed by immunoblotting. Cells were and stimulated with 1 µg/ml LPS for the indicated time and cell lysates were immunoblotted for pERK, ERK, and GAPDH. (B) Wild type (WT) or DOK3-deficient BMM were stimulated with 1 µg/ml LPS and cell lysates were immunblotted for pERK, ERK, and the GAPDH. The amount of DOK3 and Grb2 was detected by immunoblot in cell lysates. (C) BMM cells were pretreated with DMSO or 10 µM MG132 and stimulated with 1 µg/ml LPS for the indicated times. Lysates were immunoblotted for phosphorylated ERK (pERK), ERK, and DOK3. (D) RAW264.7 macrophages transfected with FLAG-tagged mouse DOK3- were stimulated with 1 µg/ml LPS for the indicated time and cell lysates were immunoblotted for pERK, ERK, FLAG and GAPDH. Data are representative of three independent experiments.