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. Author manuscript; available in PMC: 2019 Sep 12.
Published in final edited form as: Mol Pharm. 2019 May 13;16(6):2354–2363. doi: 10.1021/acs.molpharmaceut.8b01274

Figure 2.

Figure 2.

Fluorescence values of anti-Early Endosomal Antigen-1 (EEA1) staining in CEACAM1 HeLa cells as determined using internalization adaptive erode masks. (A) CEACAM1 HeLa cells were dissociated, fixed with 4% PFA, and stained with an αEEA1 antibody (green). Internalization masks were defined as being a set percent of the full brightfield mask (75–95%) while surface masks were defined as being fluorescence outside of a set internalization mask. Masks were used to determine adhered versus internalized fluorescence. White scale bar is set to 7 μm. (B) When varying internalization and surface masks are used to quantify EEA1 fluorescence, which should only be within cells, fluorescence was considered internalized and not surface as long as surface masks were set to quantify fluorescence outside of at least the 90% Adaptive Erode internal mask. Error bars represent 95% C.I.