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. 2018 Jan 15;19(3):e44767. doi: 10.15252/embr.201744767

Figure EV5. ERRFI1 regulates AKT‐PHLPP interaction.

Figure EV5

  1. MDA‐MB‐468, PANC1, U251, and HCT116 cells were serum‐starved for 36 h and then treated with EGF (100 ng/ml) for 30 min prior to cell lysis. Cells lysates were subjected to immunoprecipitation with control IgG or anti‐ERRFI1 antibody. The immunoprecipitates were blotted with the indicated antibodies. The ERRFI1‐AKT and ERRFI1‐EGFR interaction was quantified using ImageJ. The immunoprecipitated ERRFI1 levels were normalized for each interaction and then corrected for MDA‐MB‐468 no EGF treatment. The interaction in no EGF‐treated (‐EGF) MDA‐MB‐468 is set to 1. Error bars represent ± SEM of three independent experiments. The significant difference between vehicle and EGF treatment is indicated by: **P < 0.01. Statistical analyses were performed with Student's t‐test.
  2. U251 and HCT116 cells were transfected with ERRFI1 construct. After 48 h, cells were lysed and subjected to immunoprecipitation with control IgG or anti‐PHLPP antibody. The immunoprecipitates were then blotted with the indicated antibodies.
  3. U251 and HCT116 cells from (B) were treated with increasing dose of gemcitabine (Gem) alone or in combination with 10 μM MK‐2206 2HCl (MK) for 72 h. Cell survival was then determined. Each point shows the mean values for three independent experiments; error bars represent ± SEM.

Source data are available online for this figure.