4.2. Cell Lysis and Western Blot Analysis

EJ Eva Jiménez-Mora
BG Beatriz Gallego
SD Sergio Díaz-Gago
ML Marina Lasa
PB Pablo Baquero
AC Antonio Chiloeches
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Total cell extracts preparation and Western blot analysis were performed as previously described [13]. The antibodies used were anti-BRAF and anti-AMPK (Santa Cruz Biotechnology, Dallas, TX, USA); anti-p62 (BD Biosciences, Franklin Lakes, NJ, USA); anti-ATG5, anti-phospho-LKB1 (S428), anti-LKB1, anti-phospho-ACC (S79), anti-phospho-AMPK (T172), anti-phosho-S6 (S235/236) and anti-phospho-ULK (S555) (Cell Signalling Technology, Danvers, MA, USA); anti-LC3, anti-phospho-ERK and anti-β-tubulin (Sigma, St. Louis, MO, USA); and peroxidase-conjugated secondary antibodies (DAKO, Glostrup, Denmark). For LC3-II/LC3-I ratio calculation, data densitometry of blots from different experiments was used to quantify the protein bands.

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