Staufen is a member of the family of double-stranded RNA (dsRNA)-binding proteins involved in the transport and/or localization of mRNAs to different subcellular compartments and/or organelles. These proteins are characterized by the presence of multiple dsRNA-binding domains which are required to bind RNAs having double-stranded secondary structures. The human homologue of staufen encoded by STAU, in addition contains a microtubule- binding domain similar to that of microtubule-associated protein 1B, and binds tubulin. The STAU gene product has been shown to be present in the cytoplasm in association with the rough endoplasmic reticulum (RER), implicating this protein in the transport of mRNA via the microtubule network to the RER, the site of translation.

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Western blotting analysis using STAU1 antibody . Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with STAU1 antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using NaQ™ ECL Substrate Kit . STAU1, staufen double-stranded RNA binding protein 1.

Western blotting analysis using STAU1 antibody . STAU1 expression in wild type (WT) and STAU1 shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with STAU1 antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using NaQ™ ECL Substrate Kit .

Validation of STAU1 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with STAU1 antibody and analyzed using BD flow cytometer.

Immunocytochemical staining of Hela cells using STAU1 antibody , Top panel: wild-type (WT); Bottom panal: STAU1 shRNA knockdown (KD). Nuclei were stained blue with DAPI;STAU1 was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm. Permeabilization: Triton.

Flow cytometric analysis of STAU1 expression in HT-1080 cells using STAU1antibody . Green, isotype control; red, STAU1.

Immunocytochemical staining of HT-1080 cells with STAU1 antibody . Nuclei were stained blue with DAPI; STAU1 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.

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