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

Figure 3. ERRFI1 regulates AKT‐PHLPP interaction.

Figure 3

  1. MDA‐MB‐468, PANC1, U251, and HCT116 cell lysates were subjected to immunoprecipitation with control IgG or anti‐ERRFI1 antibody. The immunoprecipitates were then blotted with the indicated antibodies. Quantifications of the Western blots were analyzed by ImageJ. To quantify each interaction, the amount of AKT or EGFR was first normalized back to the input level for each protein and then corrected by the amount of IPed protein, ERRFI1. Error bars represent ± SEM of three independent experiments. The significant difference between ERRFI1‐EGFR binding and ERRFI1‐AKT binding is indicated by: **P < 0.01. Statistical analyses were performed with Student's t‐test.
  2. Purified recombinant GST, GST‐AKT, and His‐ERRFI1 were incubated in cell‐free conditions. The interaction between AKT and ERRFI1 was then examined.
  3. MDA‐MB‐468, PANC1, U251, and HCT116 cell lysates were subjected to immunoprecipitation with control IgG or anti‐PHLPP antibody. The immunoprecipitates were blotted with the indicated antibodies. The interaction was then quantified in each cell line. The amount of AKT corrected by IPed PHLPP was calculated in each of the four cancer cells transfected with siNeg or siERRFI1s. The results were then corrected by the siNeg. The interaction in siNeg is set to 1 within each cell line. Error bars represent ± SEM of three independent experiments. The significant difference between siNeg and siERRFI1 is indicated by: **P < 0.01. Statistical analyses were performed with Student's t‐test.

Source data are available online for this figure.