Expression levels of PTEN, Phospho-ERK1/2 (P-Erk), NFκB and STAT3 were measured by immunofluorescence with specific antibodies: anti-PTEN, anti-Phospho-ERK1/2, anti-NFκB p65, anti-STAT3 in MCF-10A/Zeo and MCF-10A/ERα36 cells. Total, nuclear and cytoplasmic average fluorescent signal intensities (AlexaFluor 555) are quantified from 5 cells in at least 5 separate fields per condition. Each bar represents mean ± S.E.M. N = 3 independent experiments. *: P<0.05, ***: P <0.001. A. Average fluorescence of PTEN, Phospho-ERK1/2, NFκB and STAT3 in whole MCF-10A/Zeo and MCF-10A/ERα36 cells. B. Ratios of nuclear or cytoplasmic average fluorescence of STAT3 and NFkB intensities in MCF-10A/ERα36 versus MCF-10A/Zeo cells indicating nuclear relocalization following ERα36 overexpression.