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. Author manuscript; available in PMC: 2023 Sep 22.
Published in final edited form as: Biosci Trends. 2023 Mar 9;17(2):126–135. doi: 10.5582/bst.2023.01022

Figure 5. Recombinant ERp57 restored the effects of PGRN and ND7 on lysosomal storage in ERp57 KO L444P fibroblasts.

Figure 5.

(A) The characterization of recombinant ERp57 (rERp57) by Coomassie blue staining. (B) The endocytosis of rERp57. L444P fibroblasts were treated with His-tagged rERp57 (2 μg/mL) for 8 hours. The endocytosis of rERp57 was analyzed by immunofluorescence staining with antibody against His-probe. PBS was used as control. DAPI was used to stain the nuclei. (C) The lysosomal storage in ERp57 KO L444P. ERp57 KO L444P fibroblasts were treated with ND7 (5 μg/mL), or PGRN (10 μg/mL), or ND7/PGRN plus rERp57 (0.5 μg/mL or 2 μg/mL) for 24 hours. PBS and ERp57 (2 μg/mL) were used as controls. The fluorescence intensity of the LysoTracker Red was read using SpectraMax i3x plate reader at excitation/emission of 647/668 nm. (D) The live fluorescence imaging of lysosomal storage in ERp57 KO L444P fibroblasts with or without PGRN (10 μg/mL), ND7 (5 μg/mL), PGRN plus rERp57 (2 μg/mL), or ND7 plus rERp57 (2 μg/mL), measured by LysoTracker Red. PBS used as control. (E) Quantification of mean fluorescence intensity of D. Data are shown as mean ± SD of 3 independent experiments. One-way ANOVA tests. *, p<0.05, **, p<0.01. ns: not significant. Scale bar=100 μm.