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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 1: The effects of PGRN and ND7 on GD fibroblasts is Hsp70-independent.

Figure 1:

(A) The protein level of Hsp70 in mouse lung epithelium cells, assayed by western blotting. (B) Diagram of PGRN and ND7 (the 7th N-terminal deletion). PGRN regions were highlighted in yellow, which include seven and half granulin unit (P, G, F, B, A, C, D, E). The full regions linkers were highlighted in green, and half region (P) linker was highlighted in light blue, respectively. (C) Expression and characterization of recombinant His tagged-PGRN. His-tagged PGRN protein was purified from the HEK293 cells expressing His-PGRN using His-Select Nickel Affinity Gel. Purified PGRN was analyzed by Coomassie blue staining (left) and western blotting with PGRN antibody (right). (D) Expression and characterization of recombinant His tagged-ND7. Purified ND7 was analyzed by Coomassie blue staining (left) and western blotting with PGRN antibody (right). (E) PGRN and ND7 effectively reduce lysosomal storage in Hsp70 knockout cells. Hsp70 KO lung epithelium cells were stimulated with lipid lysis and treated with recombinant ND7 (5 μg/mL) or PGRN (10 μg/mL) for 24 h, PBS served as control. The cells were stained with LysoTracker Red. The live fluorescence microscopy imaging was taken using fluorescence microscope. (F) The quantification analysis of the mean florescence intensity E. Data are shown as mean ± SD of 3 independent experiments. One-way ANOVA tests. *, p<0.05, **, p<0.01. Scale bar=100 μm.