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. 2023 Aug 11;30(10):2351–2363. doi: 10.1038/s41418-023-01205-1

Fig. 1. Identification of BNIP3 and BNIP3L proteins as FBXL4 interacting proteins.

Fig. 1

A Tandem affinity purification of FBXL4-containing protein complexes was conducted from 293 T cells stably overexpressing FLAG-FBXL4. The number of total/unique peptides identified by mass spectrometry analysis are shown in the Table. B, C 293 T cells were transfected with the indicated plasmids. The whole cell lysates (WCL) were prepared and subjected to co-IP with anti-FLAG antibody. The immunoprecipitates were analyzed by WB with the indicated antibodies. D Recombinant expressed GST-FBXL4 protein or GST bound to glutathione-Sepharose beads and incubated with recombinant expressed His-BNIP3 or BNIP3L proteins. Bound His-BNIP3 or BNIP3L proteins were detected by WB with anti-His antibody. E Schematic representation of FBXL4 deletion mutants. F 293 T cells were transfected with the indicated plasmids. The WCL were prepared and subjected to co-IP with anti-FLAG antibody. The immunoprecipitates were analyzed by WB with the indicated antibodies.