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. 2014 Feb 19;289(14):10045–10056. doi: 10.1074/jbc.M113.521880

FIGURE 5.

FIGURE 5.

FLJ00018 is activated through Ras/MAPK pathway. A and D, NIH3T3 cells were co-transfected with expression vectors for FLJ00018 (F018) and EGFR as indicated. Transfected cells were treated with 10 μm KN-93 (A), 10 μm U-0126 (A), 10 μm SB202190 (SB) (D), or 10 μm SP600125 (SP) (D) for 30 min before stimulation with 20 ng/ml EGF for 15 min. Equal amounts of proteins were resolved by 7.5% SDS-PAGE. To detect Myc-tagged FLJ00018, immunoblotting (IB) was performed with antibodies against Myc. B, C, and E, NIH3T3 cells were co-transfected with pSRE.L-luciferase, pRL-SV40 plasmid DNAs, and expression vectors for FLJ00018 and EGFR as indicated. Transfected cells were treated with KN-93 (10 μm) (B), U-0126 (10 μm) (C), SB202190 (SB) (10 μm) (E), or SP600125 (SP) (10 μm) (E) for 1 h before stimulation with 20 ng/ml EGF for 6 h. Luciferase activity was determined by dual-luciferase reporter assay. Luciferase activity obtained with mock cells was taken as 1.0, and relative activities are shown. Values are the means ± S.D. from at least three experiments.