Fig. 3: POTEF interacts with CCT subunits through its own actin domain. | Cell Death Discovery

Fig. 3: POTEF interacts with CCT subunits through its own actin domain.

From: Primate-specific POTE-actin gene could play a role in human folliculogenesis by controlling the proliferation of granulosa cells

Fig. 3

A Co-immunoprecipitants with anti-Flag antibody were separated on SDS-PAGE and visualized using Negative Gel Stain MS Kit. The band around 120 kDa (arrowhead) was subsequently confirmed as POTEF with western blotting. Other unique bands visualized around 60 kDa (arrow) were identified with mass spectrometry (Supplementary Table 2). Two CCT subunits were confirmed as molecules interacting with POTEF using both anti-TCP-1α and anti-TCP-1θ antibodies. B–D Fluorescent image of CuO-POTEF-Flag HGrC1 cells with anti-Flag antibodies, anti-TCP-1α antibodies, or DAPI nuclear staining, respectively, without cumate treatment. E Merged image of (B), (C), and (D). F–H Fluorescent image of CuO-POTEF-Flag HGrC1 cells with anti-Flag antibodies, anti-TCP-1α antibodies, or DAPI nuclear staining, respectively, after 3 days in the presence of cumate solution. I Merged image of (F), (G), and (H). Scale bars, 50 µm. J Schematic comparison of human POTEF based on the immunoprecipitation experiments. Full-length POTEF has a cysteine-rich domain (CRD, orange), ankyrin repeat (Ankyrin, blue), helices domain (helices, green), actin domain (Actin, red), and Flag tag at the C-terminus. The ankyrin repeat consisted of full-length POTEF without the actin domain. The actin domain consisted of only the actin domain. K Immunoprecipitation with anti-Flag antibody followed by western blotting with anti-Flag antibody. L Co-Immunoprecipitation with anti-Flag antibody followed by western blotting with anti-TCP-1α and anti-TCP-1θ antibodies.

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