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. 2015 Dec 17;5:18465. doi: 10.1038/srep18465

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Copyright © 2015, Macmillan Publishers Limited

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Trafficking of lactoferrin by sGAPDH involves internalization via cell surface uPAR and macropinocytosis (a) Lf internalization via GAPDH is decreased in uPAR knock down cells. PMA activated wild type and uPAR knock down THP-1 cells (5 × 10⁵/aliquot) were allowed to internalize Lf-FITC (5 μg) pre-incubated with GAPDH (10 μg) in 200 μl FACS buffer for 1 hr at 37 °C. Cells were washed and treated with pronase (0.1%). Internalized Lf was quantified from 10⁴ cells by flow cytometry and presented as histogram overlay, p ≤ 0.0001. Results were verified by three repetitions of the experiment. (b) GAPDH additionally internalizes lactoferrin via the macropinocytosis pathway as demonstrated by co-localization studies in J774 cells. Arrows indicate the strong co-localization of Lf, GAPDH and dextran. Experiment was repeated thrice and representative photograph is presented.