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. 2022 Jun 3;12:862806. doi: 10.3389/fonc.2022.862806

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Copyright © 2022 Conciatori, Salvati, Ciuffreda, Shirasawa, Falcone, Cognetti, Ferretti, Zeuli, Del Bufalo, Bazzichetto and Milella

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Evaluation of PI3K pathway activation in response to Geda in the presence of fibroblast-derived CM. (A) HCT1116 Parental and HCT116 PTEN^-/- cell lines were treated with 1 and 10 nM of Geda in serum-free condition and HFF-CM. Cells were lysed and analyzed by Western Blotting using specific antibodies (as indicated). GAPDH is shown as protein loading and blotting control. (B) HCT116 Parental cells were grown in wo FBS and HFF CM for 24 h. Endogenous mTOR was immunoprecipitated (IP:mTOR) and the immunocomplexes were blotted for RAPTOR and PRAS40. mTOR, RAPTOR and PRAS40 levels in total cell lysates are also shown. Results of a representative experiment out of three independent experiments performed are shown.