Fig. 2

Trehalose induces a dose- and time-dependent increase in PGRN expression in cultured cells. a Immunoblot of cell lysates from H4 cells treated with 0, 25, 50, or 100 mM trehalose for 20 h. Quantification of b PGRN, c LC3-II, and d p-4EBP1 immunoreactivty from a (n = 3 independent experiments). e Immunoblot of cell lysates from H4 cells treated with 100 mM trehalose for 0, 8, 16, or 24 h. Quantification of f PGRN, g LC3-II, and h p-4EBP1 immunoreactivity from e (n = 3 independent experiments). i Representative image of H4 cells treated with vehicle or trehalose (100 mM) for 24 h followed by staining with an antibody against human PGRN (red). Nuclei (blue) are stained with DAPI, 4’,6-diamidino-2-phenylindole. Scale bar, 10 μm. j Immunoblot of conditioned media from H4 cells treated with 0, 50, or 100 mM trehalose for 20 h. k Quantification of secreted PGRN immunoreactivity from Western blots of conditioned media in j (n = 3 independent experiments). In all graphs, the bars represent the mean ± SEM. *Differs from control, P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 using one-way ANOVA followed by Dunnett’s comparison post-hoc test