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. 2016 Apr 29;7(23):34371–34383. doi: 10.18632/oncotarget.9110

Figure 4. FRA1 is required for AKT-activation, and promotes cancer cell growth through AKT but not c-Jun.

Figure 4

(A) FaDu cells transfected with siCon, sic-Jun or siFRA1 oligonucleotides were seeded in triplicates for 48 h growth analysis. Graph represents average percentage of cell numbers normalized to control cells + SD. (B–E) Immunoblotting of protein lysates collected from (B) FaDu cells transfected as in (A), (C) FaDu cells that had undergone 48 h serum-starvation and then time-course stimulation with 25 ng/ml EGF, (D) FaDu and SCC25 cells transfected with siCon or siFRA1 oligonucleotides, and (E) FaDu cells that had been treated with JNK/c-Jun(SP600125), MEK/ERK(PD98059), PI3K/AKT(LY294002) inhibitors for 24 h. Relative densitometry was shown below each band. (F) Soft agar colony assay. FaDu cells transduced with lentiviruses for expression of shCon or shFRA1 either with or without the active form of AKT1. Graph represents average percentage of colonies normalized to shCon group + SD. Gene silencing and overexpression were confirmed by immunoblotting as shown on the right panel.