Granulins are a family of secreted, glycosylated peptides that are cleaved from a single precursor protein with 7.5 repeats of a highly conserved 12-cysteine granulin/epithelin motif. The 88 kDa precursor protein, progranulin, is also called proepithelin and PC cell-derived growth factor. Cleavage of the signal peptide produces mature granulin which can be further cleaved into a variety of active, 6 kDa peptides. These smaller cleavage products are named granulin A, granulin B, granulin C, etc. Epithelins 1 and 2 are synonymous with granulins A and B, respectively. Both the peptides and intact granulin protein regulate cell growth. However, different members of the granulin protein family may act as inhibitors, stimulators, or have dual actions on cell growth. Granulin family members are important in normal development, wound healing, and tumorigenesis.

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Flow cytometric analysis of Granulin precursor expression in HepG2 cells using Granulin precursor antibody . Green, isotype control; red, Granulin precursor.

Immunocytochemical staining of HepG2 cells with Granulin precursor antibody . Nuclei were stained blue with DAPI; Granulin precursor was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Low. Scale bar, 20 μm.

Western blotting analysis using granulin precursor antibody . Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with granulin precursor antibody and HRP-conjugated goat anti-mouse secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Western blotting analysis using granulin precursor antibody . Granulin precursor expression in wild type (WT) and granulin precursor (GRN) shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with granulin precursor antibody and HRP-conjugated goat anti-mouse secondary antibody respectively. Image was developed using NaQ™ ECL Substrate Kit .

Validation of GRN knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with GRN antibody and analyzed using BD flow cytometer.

Immunocytochemical staining of HeLa cells using Granulin precursor antibody , Top panel: wild-type (WT); Bottom panal: Granulin precursor shRNA knockdown (KD). Nuclei were stained blue with DAPI; Granulin precursor was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.

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