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. Author manuscript; available in PMC: 2013 Aug 1.
Published in final edited form as: FEBS J. 2012 Jul 2;279(15):2695–2713. doi: 10.1111/j.1742-4658.2012.08652.x

Figure 9. Inhibition of M6P/IGF2R Ectodomain Shedding by 1,10-Phenanthroline.

Figure 9

HEK 293 cells were transfected with empty pCMV5 vector (25 μg DNA) (A) or with the cDNA constructs indicated along the right side of each panel: B) 25 μg WT-M cDNA; C) 25 μg Rep9F cDNA; or D) 25 μg Ref14F cDNA. On the day after transfection, the cells were switched to serum-free medium plus the indicated concentrations of 1,10-phenanthroline. On the third day after transfection, 48-h concentrated conditioned medium (CCM) and whole-cell lysates were prepared. Aliquots (100 μg protein) of the lysates and CCM (50 μl volume) were analyzed by immunoblotting with the antibodies indicated to the right of each panel. The blots for actin as loading control apply to lysate samples only. These data are representative of three replicate experiments. Regression analysis indicated that the effects of OPA treatment on retention of the various receptor species in lysates were significant: A) P <0.005; B) P <0.02; C) P <0.006; D) P <0.005.