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4.8. Protein Extraction and Immunoblotting

FB Fatima Bouhtit
MN Mehdi Najar
DA Douâa Moussa Agha
RM Rahma Melki
MN Mustapha Najimi
KS Khalid Sadki
NB Noureddine Boukhatem
DB Dominique Bron
NM Nathalie Meuleman
AH Abdellah Hamal
LL Laurence Lagneaux
PL Philippe Lewalle
MM Makram Merimi
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Cells were seeded in 72 cm2 at a density of 0.8 × 106 cells/mL. After treatment with the indicated concentrations of the specified drugs for the indicated time points, cells were harvested and washed twice with PBS. Total protein extraction was performed by using the M-PER Mammalian Extraction Reagent from Thermo Fisher (78505) supplemented phosphatase/protease inhibitor cocktail from Thermo Fisher (78440). The protein concentration was determined by BCA Protein Assay (23252). Western blotting was performed as described previously [36]. Caspase-3 (#9668), caspase-9 (#9508), CHOP (#2895), p-AKT (#4060), and PI3K antibodies were purchased from Cell Signaling.

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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 (http://creativecommons.org/licenses/by/4.0/).

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