Fig. 6

SPRY2 suppresses the Y10 phosphorylation of LDHA by interfering LDHA and SRC interaction. A GSEA plot based on the gene expression profiles of SPRY2-low group compared with SPRY2-high group in GEO data sets GSE9014. NES, normalized enrichment score. False discovery rate (FDR) was set at 0.25. B ERK total protein expression and ERK phosphorylation level was determined by western blot after SPRY2 knockdown in fibroblasts. C Concentration of lactate in medium after treated with ERK signaling inhibitor U0126 in fibroblasts with SPRY2 knockdown. D LDHA total protein expression and LDHA Y10 phosphorylation level was determined by western blot after SPRY2 knockdown in fibroblasts. E LDHA enzyme activity in SPRY2 knockdown or overexpressing cells. F Co-IP analysis of the interaction between SRC and LDHA in fibroblasts transfected with vector (NC) or HA-Spry2 expressing plasmids. G Fibroblasts transfected with HA-Spry2 expressing plasmids were lysed and immunoprecipitated with anti-IgG or anti-HA antibody and immunoblotted with the indicated antibodies. H Concentration of lactate in medium of fibroblasts treated with or without SRC inhibitor KX2-391 in fibroblasts with SPRY2 knockdown or not. Data are presented as mean ± SD (n = 3 independent samples). I Western blot showing protein expression of p-SRC, total SRC, p-LDHA and LDHA in fibroblasts treated with or without SRC inhibitor KX2-391 in fibroblasts with SPRY2 knockdown or not. P values based on unpaired Student’s t test: *P < 0.05, **P < 0.01, ***P < 0.001