Fig. 1: STAT5A interacts with PDC in 293T cells. | Cell Death & Disease

Fig. 1: STAT5A interacts with PDC in 293T cells.

From: Mitochondrial STAT5A promotes metabolic remodeling and the Warburg effect by inactivating the pyruvate dehydrogenase complex

Fig. 1: STAT5A interacts with PDC in 293T cells.

a, b Tandem affinity purification of STAT5A-containing protein complexes was conducted using 293T cells stably expressing FLAG-HA-STAT5A. Associated proteins were separated by SDS-PAGE and visualized by Coomassie Blue (CB) staining (a). The numbers of total/unique peptides identified by mass spectrometry are shown in the table (b). c 293T cells were co-transfected with indicated plasmids. Whole-cell lysates (WCL) were immunoprecipitated with anti-FLAG antibody, and the immunoprecipitated proteins were immunoblotted with antibodies specific to the Myc or the FLAG. The total proteins used for immunoprecipitation were 4–5 mg. The proteins used for Immunoblot were 20–30 μg per lane. d Co-immunoprecipitation of STAT5A with PDC from 293T cells. WCL (lane 1) or immunoprecipitates generated with the STAT5A antibody (lane 3) or a control IgG (lane 2) were immunoblotted with indicated antibodies. The total proteins used for immunoprecipitation were 10–12 mg. The proteins used for Immunoblot were 45–60 μg per lane. e Co-immunoprecipitation of PDHA1 with STAT5A from 293T cells. WCL (lane 1) and immunoprecipitates generated with the PDHA1 antibody (lane 3) or a control IgG (lane 2) were immunoblotted with indicated antibodies. The total proteins used for immunoprecipitation were 10–12 mg. The proteins used for immunoblot were 45–60 μg per lane. f 293T cells were co-transfected with indicated expression vectors. WCL were immunoprecipitated with anti-FLAG antibody, and the immunoprecipitated proteins were immunoblotted with antibody specific to the Myc (for STAT5A; upper panel) or the FLAG (lower panel). The total proteins used for immunoprecipitation were 4–5 mg. The proteins used for immunoblot were 20–30 μg per lane. g 293T cells were transfected with expression vectors encoding FLAG-STATs. WCL were immunoprecipitated with anti-FLAG antibody, and the immunoprecipitated proteins were immunoblotted with indicated antibodies. The total proteins used for immunoprecipitation were 4–5 mg. The proteins used for Immunoblot were 20–30 μg per lane.

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