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Fig. 1 | Cellular & Molecular Biology Letters

Fig. 1

From: Esterase D stabilizes FKBP25 to suppress mTORC1

Fig. 1

ESD interacted with N-terminus domain of FKBP25 (1–90 aa). a, b Co-immunoprecipitation (Co-IP) of ESD with the FKBP25 protein or ectopically expressed GFP-FKBP25 from HEK293T cells. c Double immunofluorescence staining of GFP-ESD (green) and FKBP25 (red) showed agglomeration and co-localization in HeLa cells. Overlay of the GFP-ESD and FKBP25 with yellow indicates co-localization. Scale bar: 25 μm. d, e A549 cell line was transfected with plasmid pEGFP-FKBP25 for 24 h and then exposed to FPD5 (5 μM) for 24 h. Co-IP showed that ESD could bind more FKBP25 induced by FPD5. f, g HEK293T cell line was transfected with plasmids pCAG-ESD-WT and pCAG-ESD-K213A for 48 h. Then western blot analysis of co-immunoprecipitation of RFP-ESD with anti-FKBP25 antibodies and quantification of RFP-ESD level. h HEK293T cell line was transfected with plasmids FKBP25-WT and the deletion mutants of FKBP25 for 24 h respectively. Then the levels of GFP-FKBP25 and ESD were analyzed after Co-IP of ESD with anti-GFP antibodies. Data are mean ± SEM. *p < 0.05, **p < 0.01, n = 3

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