Fig. 2
From: Activation of MEK2 is sufficient to induce skin papilloma formation in transgenic zebrafish
Constitutively activated MEK1 (MEK1S219D) and MEK2 (MEK2S219D) both phosphorylated ERK1. a. COS-1 cells were transiently transfected with pcDNA3-HA, pcDNA3-ERK1-HA, pcDNA3-MEK1-mCherry, pcDNA3-MEK1S219D-GFP, pcDNA3-MEK2-GFP, or pcDNA3-MEK2S219D-GFP. Total lysates were analyzed by Western blotting using anti-HA, anti-GFP, anti-pERK monoclonal antibodies; anti-mCherry and anti-Actin polyclonal antibodies. b. Intracellular localization of ERK1, MEK1, and MEK2 in COS-1 cells by fluorescent microscopy. The pcDNA3-ERK1-HA was co-transfected with pcDNA3-GFP (A1-A4), pcDNA3-MEK1-mCherry (B1-B4), pcDNA3-MEK1S219D-GFP (C1-C4), pcDNA3-MEK2-GFP (D1-D4), or pcDNA3-MEK2S219D-GFP (E1-E4). Cy2 or Cy3 dye used an anti-HA monoclonal antibody to detect localization of ERK1 as visualized. DAPI was used to stain nuclear DNA. White arrows indicate the ERK1 protein localized in nuclei and the cytoplasm. IB, immunoblot