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. 2023 Apr 14;24(8):7273. doi: 10.3390/ijms24087273

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© 2023 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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ELISA analysis of IGF-1 secretion before and after treatment of HepG2 cells with CM from PHT cells with the control, RAPTOR, and RICTOR siRNA and densitometric analysis of mTORC1 and mTORC2 functional readouts from cell lysates of HepG2 treated with CM from PHT cells with the control, RAPTOR, and RICTOR siRNA. IGF-1 secretion was determined in CM from PHT cells with the control, RAPTOR, and RICTOR siRNA before treatment of HepG2 cells (A) and CM from HepG2 cells after treatment with CM from PHT cells with the control, RAPTOR, and RICTOR (B). Equal aliquots (50 μL) of CM were analyzed with an IGF-1 ELISA kit. A representative Western blot of HepG2 cell lysates (35 μg protein) displaying p-P70S6K ^Thr389 (C) and p-Akt ^Ser473 (D) in HepG2 cells treated with CM from PHT cells with the control, RAPTOR, or RICTOR siRNA. Samples were normalized to the control. The data were represented as Mean + SEM in bar figures and analyzed with a one-way ANOVA and Bonferroni Multiple Comparison tests; p < 0.05 was considered significant.