To assess whether PlexinD1 influences Gli1 nuclear translocation, a mechanism reported for modulation of Gli1 transcriptional activity in response to mitogenic signaling in cancer (Stecca et al, 2007), we performed a double immunofluorescence (IF) assay to visualize the nuclear localization of Gli1 in the PlexinD1’s context using α-tubulin as a cytoplasm marker. This evidence concerns the gene GLI1 and cancer.