Protein Expression Analysis via Western Blot

Western blot analyses were performed using 25 ~ 50 μg of cleared cell lysates as described previously28 (link). Antibodies used in this study were purchased from the following sources: phospho-AKT (S473) (#9271), AKT (#9272), phospho-ERK1/2 (T202/Y204) (#4370), and HER2 (#2247) from Cell Signaling Technologies, Inc. (Danvers, MA); HO-1 (ADI-OSA-110) from Enzo Life Sciences (Farmingdale, NY); β-actin, ERK1 (sc-94), NRF2 (sc-13032), and MRP5 (sc-5770) from Santa Cruz Biotechnology (Santa Cruz, CA); FLAG M2 (F1804) from Sigma; and GFP (ab290-50) from Abcam (Cambridge, MA). Chemiluminescence reagent was purchased from Santa Cruz Biotechnology or Thermo Scientific (Rockford, IL). Immunoprecipitation was performed as described previously16 (link).

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Kang H.J., Yi Y.W., Hong Y.B., Kim H.J., Jang Y.J., Seong Y.S, & Bae I. (2014). HER2 confers drug resistance of human breast cancer cells through activation of NRF2 by direct interaction. Scientific Reports, 4, 7201.

Publication 2014

HO-1 (ADI-OSA-110) β-actin FLAG M2 (F1804)

Actin Antibodies Cell Chemiluminescence Erk1 Her2 Immunoprecipitation Nrf2 Western blot analyses

Corresponding Organization :

Other organizations : Georgetown University, Georgetown University Medical Center, Dankook University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Phosphorylation of AKT (S473)
Phosphorylation of ERK1/2 (T202/Y204)
Expression of HER2
Expression of HO-1
Expression of NRF2
Expression of MRP5

control variables

Cell lysate amount (25 ~ 50 μg)
β-actin as loading control
ERK1 as loading control

controls

Positive control: Not specified
Negative control: Not specified

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