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Fig. 9. Long-term treatment (50 minutes) with CI-976 prevents subsequent uptake of Tf from the cell surface. Clone 9 cells were treated for 50 minutes at 37°C with DMSO as a solvent control (A-C) or 20 µM CI-976 (D-F). Alexa568-Tf (A,D) was then added for 5 minutes at 37°C. Cells were fixed and stained for immunofluorescence localization of M6PRs (B,E). These results are consistent with the conclusion that CI-976 inhibits recycling of TfRs back to the cell surface.