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. 2006 Dec 21;401(Pt 2):559–567. doi: 10.1042/BJ20061081

Figure 2. The ERK pathway is not necessary or sufficient for ATF3 induction.

Figure 2

(A) HeLa cells were pretreated with DMSO or 25 μM PD98059 (PD) for 30 min, followed by anisomycin or buffer treatment (i.e. control) for 1 h, and analysed by RT–PCR with primers specific to ATF3 or GAPDH (top two panels). The effectiveness of the PD98059 pretreatment was determined by the analysis of phospho-ERK at 30 min after anisomycin treatment: immunoblot with antibody against p-ERK or ERK (bottom two panels). (B) COS-1 cells were transfected with DNA expressing β-Gal or MEK1–ERK2 for 36 h, treated with anisomycin or buffer (–) for 1 h and analysed by real-time RT–PCR. ATF3 mRNA level in the β-Gal-transfected untreated cells was arbitrarily defined as 1. Results shown represent the means±S.E.M. for three experiments. (C) The specificity of the MEK1–ERK2 construct to activate the ERK but not the JNK or p38 pathway was demonstrated by phospho-kinase blot. COS-1 cells were transfected with DNA expressing β-Gal or MEK1–ERK2 for 36 h and analysed by immunoblot with the indicated antibodies. The dotted line indicates the grouping of images from different parts of the same gel.