Fig. 6

NDUFV1 suppresses the Y10 phosphorylation of LDHA by Src. a LDHA enzyme activity in MDA-MB-231 cells overexpressing CYTL1 was detected using kit assay. b LDHA enzyme activity in CYTL1 KO cells. c LDHA enzyme activity in MDA-MB-231 cells overexpressing NDUFV1. d, e LDHA Y10 phosphorylation level was determined by western blot in the indicated cells transfected with d Flag-NDUFV1 expressing plasmids or e the shRNAs against NDUFV1. GAPDH was used as a loading control. f MDA-MB-231 orthotopic breast cancer models were established as described in Fig. 4. The indicated proteins in tumor tissues were determined by western blot. g Co-IP analysis of the interaction between Flag-tagged NDUFV1 and endogenous Src in transfected HEK293T cells. h The levels of phosphorylated Src and total Src were determined by western blot in MDA-MB-231 cells overexpressing NDUFV1. i Co-IP analysis of the interaction between Src and LDHA in HEK293T cells transfected with Flag-NDUFV1 expressing plasmids. j A schematic model for the role of intracellular CYTL1 in regulating metabolic reprogramming in breast cancer. The data are shown as the mean ± SD of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001