Figure 8. Immunofluorescence analysis of pS6 in asymmetric co-cultures.

HM8 melanoma cells and fibroblasts were first cultured within adjacent compartments and subsequently allowed to interact (illustrated by arrows), forming asymmetric co-cultures before treatment with 1 μM BRAFi for 24 h (controls were not treated). The cultures were immunostained for pS6 (red) and GFP (green); cell nuclei were stained with DAPI. The staining patterns for the areas representing the distant site (labeled as “1”) and interaction front” (labeled as “2”) are shown. Scale bar, 200 μm.