Figure 2

Amino acid increase oppositely affects H-ras and K-ras activity. (A,B) Nanoclustering-FRET analysis in HEK cells co-expressing mGFP- and mCherry-tagged (A) H-rasG12V or (B) K-rasG12V. Cells were incubated for 24 h with increasing concentrations of amino acids (1x, 4x or 8x) in the absence or presence of serum (FCS) as indicated. The numbers in the bars indicate the number of analyzed cells (mean ± SEM, n = 3). (C,D) Western blot analysis of Ras- and mTORC1-signalling in HEK cells expressing mGFP-H-rasG12V or mGFP-K-rasG12V. Cells were treated as in (A,B) under serum-starved conditions. Numbers indicate the ratio of phosphorylated to respective total protein levels (n = 4). Statistical significance levels are annotated as *p < 0.05; ****p < 0.0001.