Fig. 5

Trehalose rescues PGRN deficiency in patient-derived primary fibroblasts. a Top, Immunoblot of cell lysates from primary human fibroblast (3 controls; CTL, and 3 progranulin mutation; GRN) cell lines treated with vehicle or trehalose (100 mM) for 24 h. Bottom, Immunoblot of conditioned serum-free media from primary human fibroblast cell lines treated with vehicle or trehalose (100 mM) for 20 h. b Quantification of intracellular PGRN from immunoblots in a (top). c Quantification of secreted PGRN from immunoblots in a (bottom). d GRN mRNA levels were quantified from control cell lines (CTL) or progranulin mutation cell lines treated with vehicle or trehalose (100 mM) for 18 h (each individual line run in duplicate on qPCR). e GRN primary fibroblasts treated with trehalose (100 mM, 20 h) showed increased cytoplasmic staining of PGRN (red) similar to CTL fibroblasts. Scale bar, 10 μm. Nuclei (circles) and cell bodies are outlined in white. In all graphs, the data are normalized to the lowest CTL value for comparison. For b and c, bars represent individual densitometric measurements from the immunoblots in a and for d, bars represent the mean ± SEM of two qPCR replicate values. *Differs from untreated GRN, P < 0.05, **P < 0.01, ***P < 0.001, using unpaired two-tailed Student’s t-test