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2 protocols using microtubule associated protein 1 light chain 3

1

Western Blot Analysis of Apoptosis and Autophagy

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For western blots, 2 × 105 OVCAR-3 or A2780 cells per well were plated in a 6-well plate for 24 h. Subsequently, the cells were incubated with the appropriate drug for distinct time periods and the western blots were performed as previously described [36 (link)] using the following antibodies: Capsese-3 (Cell Signaling Technology, USA, #9661), PARP (Cell Signaling Technology, USA, #9542), Microtubule-associated protein 1 light chain 3 (LC3) (Sigma, USA, #L7543), p62/SQSTM1/SQSTM1 (Cell Signaling Technology, USA, #5114), ATG5 (Cell Signaling Technology, USA, #9980), ATG7 (Cell Signaling Technology, USA, #8558), p-AKT (Cell Signaling Technology, USA, #9271), AKT (Cell Signaling Technology, USA, #9272), BAX (Proteintech Group, USA, #23931-1-AP), Bcl-2 (Proteintech Group, USA, #12789-1-AP), and Mcl-1 (Proteintech Group, USA, #16225-1-AP), p-p70S6K (Thr389) (Sigma, USA, #MABS82), p70S6K (Sigma, USA, #06-926), p-mTOR (S2448) (Abcam, USA, #ab109268), -S6 Ribosomal (Cell Signaling Technology, USA, #5364), S6 Ribosomal (Cell Signaling Technology, USA, #2317), mTOR (Abcam, USA, #ab2732), and β-actin (Origene, USA, #TA811000), peroxidase-linked anti-rabbit antibody (Cell Signaling Technology, Danvers, USA, #7074) or peroxidase-linked anti-mouse antibody (Sigma-Aldrich, St. Louis, USA, #A9044).
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2

Chemicals and Antibodies for Cell Signaling Assays

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The chemicals used in our experiments were: trichostatin A (Sigma, T8552), bafilomycin A1/BAF (Sigma, B1793), and chloroquine (CQ, Sigma, C6628). The antibodies used in our experiments included: FOXO1 (Cell Signaling Technology, 2880), phospho-FOXO1 (Cell Signaling Technology, 9461), P21 (Cell Signaling Technology, 2947), microtubule-associated protein 1 light chain 3/LC3 (Sigma, L7543), p62 (Sigma, P0067), phospho-AKT (Cell Signaling Technology, 4060), AKT (Cell Signaling Technology, 4691), phospho-S6 (S235/236; Cell Signaling Technology, 2211), S6 (Cell Signaling Technology, 2217), PARP1 (Cell Signaling Technology, 9542), caspase 3 (Cell Signaling Technology, 9662), β-actin (Sigma, A5441), α-tubulin (Sigma, T6199), and lamin AC (Cell Signaling Technology, 2032).
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