Fig. 3

Immunofluorescence characterization of 3D-hCAFs/iCCA spheroids. (A). Representative confocal microscopy images of homo- and hetero-spheroids established by KKU-M213 and KKU-M156 alone or in co-culture with hCAFs. The EpCAM marker, considered an epithelial-specific marker, is shown as a green fluorescent signal. The FAP marker, used as a specific marker for fibroblast activation, is shown as a red fluorescent signal. (B). Positive staining for Ki-67 indicates cell proliferation and demonstrates that hCAFs influence proliferation in hetero-spheroids. Nuclei are shown as a blue fluorescent signal stained with DAPI. (20× magnification, scale bar 100 μm)