The encoded protein is a homolog of yeast securin proteins, which prevent separins from promoting sister chromatid separation. It is an anaphase-promoting complex (APC) substrate that associates with a separin until activation of the APC. The gene product has transforming activity in vitro and tumorigenic activity in vivo, and the gene is highly expressed in various tumors. The gene product contains 2 PXXP motifs, which are required for its transforming and tumorigenic activities, as well as for its stimulation of basic fibroblast growth factor expression. It also contains a destruction box (D box) that is required for its degradation by the APC. The acidic C-terminal region of the encoded protein can act as a transactivation domain. The gene product is mainly a cytosolic protein, although it partially localizes in the nucleus. Three transcript variants encoding the same protein have been found for this gene.

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Immunocytochemical staining of HAP-1 cells with PTTG1 antibody . Nuclei were stained blue with DAPI; PTTG1 was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Scale bar, 20 μm.

Validation of PTTG1 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with PTTG1 antibody and analyzed using BD flow cytometer.

Flow cytometric analysis of PTTG1 expression in HAP-1 cells using PTTG1 antibody . Green, isotype control; red, PTTG1.

Western blotting analysis using PTTG1 antibody . Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with PTTG1 antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Western blotting analysis using PTTG1 antibody . PTTG1 expression in wild type (WT) and PTTG1 shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with PTTG1 antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using NaQ™ ECL Substrate Kit .

Immunocytochemical staining of HeLa cells using PTTG1 antibody , Top panel: wild-type (WT); Bottom panal: PTTG1 shRNA knockdown (KD). Nuclei were stained blue with DAPI; PTTG1 was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.

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