hek293t cells  (ATCC)

Journal: bioRxiv

Article Title: AKIP1 is an inner scaffold component required for centriole integrity

doi: 10.64898/2026.06.03.729831

Figure Lengend Snippet: (A-D) Co-immunoprecipitation assay in HEK293T cells cotransfected with Flag-AKIP1 and HA-POC1A (A), Flag-AKIP1 and HA-POC1B (B), Flag-AKIP1 and HA-Centrin-2 (D), or transfected with Flag-AKIP1 alone (C). Cell lysates were immunoprecipitated with FLAG antibodies and immunoblotted with the indicated antibodies. (E) Predicted structure and predicted aligned error plots of AKIP1 retrieved from the AlphaFold Protein Structure Database (AF-Q9NQ31-F1), colored according to pLDDT. Blue: pLDDT > 90; cyan: 90 > pLDDT > 70; yellow: 70 > pLDDT 50; orange: pLDDT < 50. (F) Schematic representation of the predicted structural organization of AKIP1 based on (E). (G-I) AlphaFold3 predictions of complexes and predicted aligned error plots of its predictions for AKIP1 and POC1A (G), POC1B (H), or POC5 (I). Red and magenta dashed boxes indicate predicted interaction interfaces. (J) Schematic of the molecular interactions among known inner scaffold factors and AKIP1. Solid black lines indicate interactions established in previous studies, cyan lines represent those identified in this study, and the dashed line indicates an interaction reported previously but currently debated , .

Article Snippet: HEK293T cells were collected 24 h after transfection and lysed on ice in lysis buffer (50 mM Tris-HCl pH 7.5, 200 mM NaCl, 0.5% Triton X-100, 1 mM DTT, and 1:500 protease inhibitor cocktail (Nacalai Tesque, 25955-11)).

Techniques: Co-Immunoprecipitation Assay, Transfection, Immunoprecipitation