FIGURE 6.

Man-6-P-modified LIF is subject to intracellular lysosomal targeting and degradation. A, HeLa cells were transfected with various LIF-mCherry or mCherry (with signal peptide) constructs and grown in the presence or absence of 5 mm Man-6-P. Cell and media (M) pools were then analyzed by Western blot using either anti-LIF or anti-mCherry antisera. IB, immunoblot. B–E, confocal analysis of HeLa cells transfected with LIF-mCherry or mCherry constructs (red) and co-stained with the lysosomal marker LAMP-1 (green). B, LIF-mCherry. Co-localization indicates lysosomal targeting of LIF (see arrowheads). C, signal peptide-containing mCherry. D, LIF-mCherry-transfected cells grown in the presence of 5 mm Man-6-P. E, TDM31 LIF-mCherry.