The SMAD family of proteins are a group of intracellular signal transducer proteins similar to the gene products of the Drosophila gene 'mothers against decapentaplegic' (Mad) and the C. elegans gene Sma. The SMAD3 protein functions in the transforming growth factor-beta signaling pathway, and transmits signals from the cell surface to the nucleus, regulating gene activity and cell proliferation. This protein forms a complex with other SMAD proteins and binds DNA, functioning both as a transcription factor and tumor suppressor. Mutations in this gene are associated with aneurysms-osteoarthritis syndrome and Loeys-Dietz Syndrome 3.

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Western blotting analysis using phospho-Smad3 (S423 + S425) antibody . Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with phospho-Smad3 (S423 + S425) antibody and HRP-conjugated goat anti rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Western blotting analysis using phospho-smad3 (S423 + S425) antibody . Phospho-smad3 (S423 + S425) expression in wild-type (WT) and SMAD3 knockout (KO) HSHC cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with phospho-smad3 (S423 + S425) antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using NaQ™ ECL Substrate Kit .

Flow cytometric analysis of Phospho-Smad3 (S423 + S425) expression in HAP-1 cells using Phospho-Smad3 (S423 + S425) antibody . Green, isotype control; red, Phospho-Smad3 (S423 + S425).

Immunocytochemical staining of HAP-1 cells with Phospho-Smad3 (S423 + S425) antibody . Nuclei were stained blue with DAPI; Phospho-Smad3 (S423 + S425) 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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