This gene encodes one member of a family of translation repressor proteins. The protein directly interacts with eukaryotic translation initiation factor 4E (eIF4E), which is a limiting component of the multisubunit complex that recruits 40S ribosomal subunits to the 5' end of mRNAs. Interaction of this protein with eIF4E inhibits complex assembly and represses translation. This protein is phosphorylated in response to various signals including UV irradiation and insulin signaling, resulting in its dissociation from eIF4E and activation of mRNA translation.

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Immunocytochemical staining of HeLa cells using Phospho-4E BP1 (Thr46) antibody , Top panel: wild-type (WT); Bottom panal: Phospho-4E BP1 (Thr46) shRNA knockdown (KD). Nuclei were stained blue with DAPI; Acyl-CoA dehydrogenase short chain was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.

Flow cytometric analysis of phospho-4E BP1 (Thr46) expression in C2C12 cells using phospho-4E BP1 (Thr46) antibody . Green, isotype control; red, phospho-4E BP1 (Thr46).

Immunocytochemical staining of C2C12 cells with Phospho-4E BP1 (Thr46) antibody . Nuclei were stained blue with DAPI; Phospho-4E BP1 (Thr46) 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.

Western blotting analysis using phospho-4E BP1 (Thr46) antibody . Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with phospho-4E BP1 (Thr46) antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Western blotting analysis using phospho-4E BP1 (Thr46) antibody . Phospho-4E BP1 (Thr46) expression in wild-type (WT) and EIF4EBP1 shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with phospho-4E BP1 (Thr46) antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using NaQ™ ECL Substrate Kit . EIF4EBP1, eukaryotic translation initiation factor 4E binding protein 1.

Validation of 4E BP1 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with Phospho-4E BP1 (Thr46) antibody and analyzed using BD flow cytometer.

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