Figure 4. Effects of SFN on Gli translocation and transcription.

(A), The nuclear translocation of Gli1 and Gli2, was measured by immunocytochemistry. Pancreatic CSCs were treated with sulforaphane (0–20 µM) for 24 h. Cells were then stained with anti-Gli and Gli2 antibody (green fluorescence), and DAPI (red fluorescence). Merged images are shown, which indicate yellow-orange staining of Gli 1 and Gli2 located in the nucleus due to co-localization of green and red fluorescence. (B), Inhibition of Gli transcription. Pancreatic CSCs were transduced with Gli-responsive GFP/firefly luciferase viral particles (pGreen Fire1-Gli with EF1, System Biosciences). After transduction, culture medium was replaced and CSCs were treated with sulforaphane (0–20 µM) for 24 h. Gli-responsive reporter activity was measured using a luciferase assay (Promega Corporation). Data represent mean ± SD. @, %, and * = significantly different from control, P < 0.05.