Western Blot Analysis of Signaling Proteins

Western blot analyses were performed as described previously (25 (link)). Antibodies used in this study were as follows: MET (sc-161), ERK1 (sc-94), and PARP (sc-7150) from Santa Cruz (Santa Cruz, CA, USA); p-EGFR (Y1068) (#2237), EGFR (#4405), p-MET (Y1234/Y1235) (#3123), phospho-AKT (Ser473) (#9271), AKT (#9272), p-ERK1/2 (T202/Y204) (#4370), p-p70 S6K (T389) (#9205), p70 S6K (#9202), p-S6 (S235/S236) (#4856), S6 (#2217) and XIAP (#2045) from Cell Signaling (Danvers, MA, USA); α-tubulin, β-actin, and horseradish peroxidase-conjugated secondary antibodies from Sigma.

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YI Y.W., YOU K., BAE E.J., KWAK S.J., SEONG Y.S, & BAE I. (2015). Dual inhibition of EGFR and MET induces synthetic lethality in triple-negative breast cancer cells through downregulation of ribosomal protein S6. International Journal of Oncology, 47(1), 122-132.

Publication 2015

PARP (sc-7150 p-EGFR (Y1068) p-MET (Y1234/Y1235) phospho-AKT (Ser473) p-ERK1/2 (T202/Y204) p-p70 S6K (T389) p70 S6K p-S6 (S235/S236) α-tubulin β-actin horseradish peroxidase-conjugated secondary antibodies

Actin Antibodies Egfr Erk1 Horseradish peroxidase P70 s6k Western blot analyses α tubulin

Corresponding Organization :

Other organizations : Dankook University

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Variable analysis

independent variables

Primary antibodies used for Western blot analysis: MET, ERK1, PARP, p-EGFR (Y1068), EGFR, p-MET (Y1234/Y1235), phospho-AKT (Ser473), AKT, p-ERK1/2 (T202/Y204), p-p70 S6K (T389), p70 S6K, p-S6 (S235/S236), S6, and XIAP

dependent variables

Protein expression and phosphorylation levels measured by Western blot analysis

control variables

α-tubulin, β-actin, and horseradish peroxidase-conjugated secondary antibodies

controls

No positive or negative controls were explicitly mentioned in the protocol.

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