Interplay between S1PR3, YAP, c-MYC and PGAM1 contributes to Warburg effect in OS. a and b. Altered level of ECAR and OCR in OS cells (MNNG-HOS and U-2OS) with treatment of Verteporfin (2 μM) or DMSO. Values are means ± SD. c and d. Interactions between c-MYC and YAP demonstrated by CO-IP assay. The input was performed as control. e. Relative mRNA levels of PGAM1 in OS cells (MNNG-HOS and U-2OS) in different groups (sh-Control, sh-S1PR3 and sh-S1PR3 + ov-YAP). Values are means ± SD, **p < .01, ***p < .001 (Student's t-test). f and g. A ChIP assay was used to verify the potential c-MYC and YAP co-binding site in the PGAM1 promoter region in MNNG-HOS and U-2OS cell lines. Input fractions and IgG were used as controls. h and i. Luciferase activities of YAP-overexpressing, c-MYC-overexpressing, YAP-overexpressing and c-MYC-knockdown or control OS cells (MNNG-HOS and U-2OS) in luciferase reporter plasmid containing wild-type and mutant 1(M1) or mutant 2 (M2) PGAM1 promoter. The data shown are mean ± SD, **p < .01, ***p < .001 (Student's t-test).