Fig. 2

TPD54 interacts with pyruvate dehydrogenase and plays an important role in breast cancer cells. a Images and the number of colony formation in control and stable TPD54 knockdown MCF-7 cells are presented. b Images and the number of colony formation of vector control and TPD54 overexpressed MCF7 cells are shown. Data were reported as the mean ± SD from triplicate data of one experiment. Values are presented as mean ± SD, student t test was used to calculate p value for two-group comparisons with ***p < 0.001 and *p < 0.05).c Endogenous TPD54 protein was detected using immunofluorescence. Mitochondria were labeled by DsRed Mito through transient transfection. The nucleus was labeled by DAPI staining. Images were taken at 200×. d Fractionations of cytoplasm (Cyto) and mitochondria (Mito) were performed in MCF-7 cells. Western blots were performed using cytoplasmic and mitochondrial fractions to identify TPD54 subcellular localization. Vinculin and NDUFB8 were used as cytoplasmic and mitochondrial markers and loading controls. e Endogenous TPD54 immunoprecipitation was performed in MCF-7 cells. Western blot detected TPD54 pulldown and its associated proteins in PDH complex. WL represents whole cell lysate as input. f PDH E1α and TPD54 were detected in MCF7 and MDA-MB-231 cells using immunofluorescence. The nucleus was labeled by DAPI staining. Images were taken at 100×. All data presented were representative experimental data that have been replicated by independent experiments