Preclinical Evaluation of Novel Apoptosis Modulators

The following drugs were kindly provided by Abbvie: venetoclax, A1210477, and A1331852. The following drugs were purchased: S63845 for in vitro and in vivo studies (S-63845; Chemietek), Lapatinib Ditosylate (Tykerb) for in vitro and in vivo studies (M1802; Abmole), and Fulvestrant (S1191; Selleckchem).
The antibodies used in this study (clone/cat. no.) were as follows: Anti-Bak (Ab-1 clone for IP) (AM03; EMD Millipore), anti-BAX (6A7 clone for IP) (sc-23959; Santa Cruz), anti-BAX (N-20 clone) (sc-493; Santa Cruz), anti-Bak (3814S; Cell Signaling), anti-Bim (C34C5) (2933S; Cell Signaling), anti-Noxa (D8L7U) (14766S; Cell Signaling), anti–Bcl-2 (D55G8) (Human Specific) (4223S; Cell Signaling), anti-Cleaved PARP (Asp214) (D64E10) (5625S; Cell Signaling), anti-GAPDH (6C5) (sc-32233; Santa Cruz), anti-HER2/ErbB2 (29D8) (2165S; Cell Signaling), anti–MCL-1 (S-19) (sc-819; Santa Cruz), anti–BCL-xL (54H6) (2764S; Cell Signaling), 4E-BP1 (53H11) (9644S; Cell Signaling), phospho-4E-BP1 (Thr37/46) (236B4) (2855S; Cell Signaling), anti–Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) (5364S; Cell Signaling), anti–Phospho-Akt (Thr308) (244F9) (4056S; Cell Signaling), anti-ERα (D8H8) (8644S; Cell Signaling), anti–Phospho-HER2/ErbB2 (Tyr1248) (2247S; Cell Signaling), anti-Akt (C67E7) (4691S; Cell Signaling), anti-cleaved Caspase-3 (Asp175) (5A1E; Cell Signaling), Normal Rabbit IgG for IP (sc-2027; Santa Cruz), and Normal Mouse IgG for IP (sc-2025; Santa Cruz).
All mouse experiments were approved and performed in accordance with the Institutional Animal Care and Use Committee at Virginia Commonwealth University (VCU).

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Floros K.V., Lochmann T.L., Hu B., Monterrubio C., Hughes M.T., Wells J.D., Morales C.B., Ghotra M.S., Costa C., Souers A.J., Boikos S.A., Leverson J.D., Tan M., Serra V., Koblinski J.E., Arribas J., Prat A., Paré L., Miller T.W., Dozmorov M.G., Harada H., Windle B.E., Scaltriti M, & Faber A.C. (2018). Coamplification of miR-4728 protects HER2-amplified breast cancers from targeted therapy. Proceedings of the National Academy of Sciences of the United States of America, 115(11), E2594-E2603.

Publication 2018

4e bp1 Anti erbb2 Antibodies Bcl 2 Caspase 3 Clone Drugs Fulvestrant Gapdh Her2 Human Institutional animal care and use committee Lapatinib ditosylate Mouse Rabbit Ribosomal protein S63845 Tykerb Venetoclax

Corresponding Organization :

Other organizations : Virginia Commonwealth University, Memorial Sloan Kettering Cancer Center, Dartmouth College, Vall d'Hebron Institut de Recerca, Centro de Investigación Biomédica en Red de Cáncer, Massachusetts General Hospital, Harvard University, AbbVie (United States), USA Mitchell Cancer Institute, University of South Alabama, Institució Catalana de Recerca i Estudis Avançats, Universitat Autònoma de Barcelona

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Variable analysis

independent variables

Venetoclax
A1210477
A1331852
S63845
Lapatinib Ditosylate (Tykerb)
Fulvestrant (S1191)

dependent variables

Cell viability
Apoptosis
Signaling pathways

control variables

Mouse experiments were approved and performed in accordance with the Institutional Animal Care and Use Committee at Virginia Commonwealth University (VCU).

controls

Positive controls: None explicitly mentioned.
Negative controls: Normal Rabbit IgG for IP, Normal Mouse IgG for IP.

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