Figure 5

Inhibition of key TCA cycle enzymes increased cell proliferation under normoxia. (A) Schema for two enzymes, citrate synthase and KGDHC, in the TCA cycle and their respective inhibitors, PCA and SP. (B) Proliferation assay of BM EPCs treated with vehicle or various concentrations of PCA under normoxia (n = 6). (C) Colony formation assay of EPCs treated with vehicle or 10uM PCA under normoxia (n = 6). (D) qRT-PCR showed higher expression of BM EPCs stemness markers in the presence of 10uM PCA than with vehicle control under normoxia (n = 5). (E) Representative experiment showing ECAR of BM EPCs and comparison of glycolytic rate and capacity in the presence or absence of PCA under normoxia (n = 6). (F) Representative experiment showing OCR of BM EPCs and comparison of basal and maximal respiration in BM EPCs in the presence or absence of PCA under normoxia (n = 6). (G) Proliferation assay of BM EPCs treated with vehicle or various concentration of SP under normoxia (n = 6). (H) Colony formation assay of BM EPCs treated with vehicle or 20uM SP under normoxia (n = 6). (I) qRT-PCR showed higher expression of BM EPCs stemness markers in the presence of 20uM SP than with vehicle control under normoxia (n = 5). (J) Representative experiment showing ECAR of BM EPCs and comparison of glycolytic rate and capacity in the presence or absence of SP under normoxia (n = 6). (K) Representative experiment showing OCR of BM EPCs and comparison of basal and maximal respiration in the presence or absence of SP under normoxia (n = 6). Data are presented as Mean ± SEM. *p < 0.05; **p < 0.01 versus vehicle control (with or without DMSO).