MEK-ERK1/2 signaling pathway is required for radiation-induced c-Jun phosphorylation.
(A) Cell lysates collected from 10 Gy-irradiated BV2 cells were immunoblotted with anti-phosphorylated MEK1/2 (sc-7995R) antibody. (B) BV2 cells were transfected with HA-MEK1, HA-MEK1K97M (kinase-dead mutant), and vector control. Protein levels of p-c-Jun and p-ERK1/2 (upper and lower panels) are shown. (C) BV2 cells that were treated or untreated with 10 μM U0126 were irradiated with 10 Gy. p-c-Jun and p-ERK1/2 levels were analyzed by Western blot. (D) Two shRNAs (sh-MEK1-212 and sh-MEK1-495) were lentivirally delivered into BV2 cells to knockdown MEK1 protein. Knockdown efficiency and protein levels of p-c-Jun and p-ERK1/2 were detected with the indicated antibodies. The quantification results for (C) and (D) are shown in Figure S2.