Fig. 3: STUB1 directly interacts with GOT2. | Cell Death & Disease

Fig. 3: STUB1 directly interacts with GOT2.

From: Glucose-induced STUB1-GOT2 axis promotes aspartate synthesis and mitochondrial dysfunction in bladder cancer

Fig. 3

A The HEK293T cells were lysed and subjected to immunoprecipitation using anti-Flag Magnetic Beads. The complexes were then separated, and the gels were stained with silver. B List of STUB1-associated proteins were identified peptides and unique peptides more than or equal three by mass spectrometric analysis. C Representative best unique peptides of STUB1 and GOT2 were identified by mass spectrometry assays. D Immunostaining of STUB1 (green) and GOT2 (red) were detected by their antibody in both UC3 and T24 cells. Nuclear 4’, 6-diamidino-2-phenylindole (DAPI; blue). E HEK293T cells were co-transfected with HA-GOT2 and Flag-tagged STUB1 plasmids, and cell lysates were subjected to IP and detected with the indicated tagged antibodies. F, G UC3 or T24 cell lysates were subjected to immunoprecipitation with control IgG, anti-STUB1 or anti-GOT2 antibody. The immunoprecipitates were then blotted with the indicated antibodies. H Purified recombinant STUB1 proteins were incubated with extracts of HA- GOT2-transfected HEK293T cells, and then immunoblotted with the antibody to HA-tag antibody. Bottom, recombinant STUB1 protein were purified from bacteria and analyzed by SDS-PAGE and coomassie blue staining. I HEK293T cells were cotransfected with HA- GOT2 plasmids and Flag-tagged full-length STUB1 plasmids or its deletion mutant plasmids for 24 h, and then cell lysates were immunoprecipitated with anti-Flag magnetic beads, and immunoblotted with antibodies to HA and Flag tag antibody.

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