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. Author manuscript; available in PMC: 2016 Nov 28.
Published in final edited form as: Cancer Discov. 2012 Aug 22;2(11):1036–1047. doi: 10.1158/2159-8290.CD-11-0348

Figure 5.

Figure 5

ERK-dependent BRCA downregulation. A, Western blot analysis of protein lysates from MDA-MB-468 cells constitutively overexpressing control or MEK1 plasmids and treated with the MEK inhibitor AZD6244 (500 nmol/L) for 4 days using the indicated antibodies. Total ERK (tERK) is used as loading control. B, qRT-PCR measuring both BRCA1 and BRCA2 mRNA levels in MDA-MB-468-MEK1 cells treated with AZD6244. Measurements were normalized to 18S mRNA levels and expressed as fold change compared to controls (log2 scale). Data are shown as mean ± SE of 3 independent replicates for each condition. C, Western blot analysis of protein lysates from MDA-MB-468 cells treated with BKM120 (750 nmol/L), AZD6244 (500 nmol/L), or the combination of both for 4 days using the indicated antibodies. Total ERK (tERK) is used as loading control. D, qRT-PCR measuring both BRCA1 and BRCA2 mRNA levels in MDA-MB-468 cells treated with BKM120 and AZD6244. Measurements were normalized to 18S mRNA levels and expressed as fold change compared with controls (log2 scale). Data are shown as mean ± SE of 3 independent replicates for each condition. E, Western blot analysis of the indicated proteins in 2 independent TNBC1 tumors treated for 4 days with BKM120 (50 mg/kg), AZD6244 (10 mg/kg), or the combination of both. Total ERK (tERK) is used as loading control. F, qRT-PCR measuring both BRCA1 and BRCA2 mRNA levels in tumor grafts. Measurements were normalized to 18S mRNA levels and expressed as fold change compared with controls (log2 scale). Data are shown as mean ± SE of 3 tumors for each condition.