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. 2015 Jul 17;10(7):e0131024. doi: 10.1371/journal.pone.0131024

Fig 1. Nuclear co-localization of p65 and RelB by a quadruple multiple staining approach.

Fig 1

A. Epithelial staining with CK18, CK19 and PSA defines the epithelial mask using orange fluorochromes (A546, Cy3). B. Identification of epithelial area (presented in grey for optimal contrast in subsequent analyses) as ROI #1 (region of interest #1). Identification of nuclei (grey surrounded by red tracing) as ROI#2 from pre-defined ROI#1. P65 and RelB fluorescence were subsequently evaluated separately in ROI#1 and #2. C. RelB staining with green fluorescent dye (A488) and p65 staining with red fluorescent dye (Cy5). Steps 1, 2 and 3 correspond to the fluorescence analysis process followed using Visiomorph DP software.