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Figure 8 | Breast Cancer Research

Figure 8

From: Nonhypoxic regulation and role of hypoxia-inducible factor 1 in aromatase inhibitor resistant breast cancer

Figure 8

Protein expression in HER2+ cells and exemestane-resistant cells. A) MCF-7Ca (M), Hc7 and LTLTCa (LT) cells were plated in their respective passage media. Total protein was extracted and HER2, phosphorylated- and total-ERK, ERα, HIF-1α, BCRP and β-actin protein were analyzed by Western blot. Shown are representative blots and overall densitometry results of n = 6 independent cell samples/group. Densitometry results are expressed as mean fold-change compared to MCF-7Ca after normalization to ERK (mean ± SD of n = 6 independent cell samples/group; *versus MCF-7Ca, P <0.05; † versus MCF-7Ca, P <0.001, one-way ANOVA). Dashed lines indicate omitted lane in between M and Hc7 of the same blots. B) AC1 (AC1) and AC1-ExR (ExR) cells were plated in their respective passage media. Total protein was extracted and HER2, phosphorylated- and total-ERK, ERα, HIF-1α and β-actin protein were analyzed by Western blot. Shown are representative blots and overall densitometry results of n = 6 independent cell samples/group. Densitometry results are expressed as mean fold-change compared to vehicle-treated cells after normalization to β-actin (mean ± SD of n = 6 independent cell samples/group; *versus vehicle, P <0.0001, two-sided t-test). C) MCF-7/HER2 cells were treated with either vehicle (V), 1 μM lapatinib (Lap), 100 nM ICI 182,780 (ICI) or 1 μM lapatinib + 100 nM ICI 182,780 (Lap + ICI) for 24 hours. Total protein was extracted and HER2, phospho- and total-ERK1/2, ERα, HIF-1α and β-actin were analyzed by Western blot. Shown are representative blots and overall densitometry results of n = 6 independent cell samples/group. Densitometry results are expressed as mean fold-change compared to vehicle-treated cells after normalization to β-actin (mean ± SD of n = 6 independent cell samples/group; *versus vehicle, P <0.0001, one-way ANOVA). ANOVA, analysis of variance; BCRP, breast cancer resistant protein; ERα, estrogen receptor alpha; HER2, human epidermal growth factor receptor 2; HIF-1α, hypoxia inducible factor 1 α subunit; n, number; SD, standard deviation.

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