Fig. 15

Immunofluorescent analysis of the DLD1Tr21/Mock and DLD1Tr21/NBPF1 cell cultures used for proteomic analysis. Cell cultures were grown for 4 days in standard medium supplemented with dox. The induced DLD1Tr21/Mock cells were post-metabolically labeled with light ¹²C₆-NHS-propionate, and the induced DLD1Tr21/NBPF1 cells were post-metabolically labeled with heavy ¹³C₆-NHS-propionate, and then submitted to proteomic analysis as described in Methods. As a control for proper NBPF1 induction, replicate cell cultures were stained for induction of EGFP and NBPF1 expression by immunofluorescent microscopy. Expression of EGFP (shown in green) is induced in both cell lines upon the addition of dox (untreated cultures were fully negative). NBPF1 (antibody sc82241; red channel) was expressed only in DLD1Tr21/NBPF1 cells and only in the presence of dox. Nuclei were visualized with DAPI. Scale bars: 10 μm