This gene belongs to the subfamily of ubiquitously expressed heterogeneous nuclear ribonucleoproteins (hnRNPs). The hnRNPs are RNA-binding proteins and they complex with heterogeneous nuclear RNA (hnRNA). These proteins are associated with pre-mRNAs in the nucleus and appear to influence pre-mRNA processing and other aspects of mRNA metabolism and transport. While all of the hnRNPs are present in the nucleus, some seem to shuttle between the nucleus and the cytoplasm. The hnRNP proteins have distinct nucleic acid binding properties. The protein encoded by this gene has four repeats of quasi-RNA recognition motif (RRM) domains that bind RNAs. This protein binds to the intronic polypyrimidine tracts that requires pre-mRNA splicing and acts via the protein degradation ubiquitin-proteasome pathway. It may also promote the binding of U2 snRNP to pre-mRNAs. This protein is localized in the nucleoplasm and it is also detected in the perinucleolar structure. Alternatively spliced transcript variants encoding different isoforms have been described.

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Immunocytochemical staining of HAP-1 cells with PTBP1 antibody . Nuclei were stained blue with DAPI; PTBP1 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.

Flow cytometric analysis of PTBP1 expression in HAP-1 cells using PTBP1 antibody . Green, isotype control; red, PTBP1.

Western blotting analysis using PTBP1 antibody . Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with PTBP1 antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Western blotting analysis using PTBP1 antibody . PTBP1 expression in wild-type (WT) and PTBP1 shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with PTBP1 antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using NaQ™ ECL Substrate Kit .

Validation of PTBP1 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with PTBP1 antibody and analyzed using BD flow cytometer.

Immunocytochemical staining of HeLa cells using PTBP1 antibody , Top panel: wild-type (WT); Bottom panal: PTBP1 shRNA knockdown (KD). Nuclei were stained blue with DAPI;PTBP1 was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm. Permeabilization: Triton.

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