Figure 6.

Quantification of total amounts by IC-MS is consistent with blockade of glutaminolysis and enhanced gluconeogenesis by ectopic miR-122 mimic expression in Gln-dependent EC4 cells. Total amounts of metabolites were quantified via IC-MS analysis as described in Methods. A-F. Depletion of selected ¹³C- and/or ¹⁵N- labeled metabolites (μmol/g protein) including glutamate (A), α-ketoglutarate (B), citrate (C), and isocitrate (D), as well as accumulation gluconeogenic products including G6P (E) and PEP (F) in EC4 cells transfected with miR122 mimic (miR122) or NC mimic (Ctl). Cx and CxNx refer to total levels of labeled products. * and ** denote Q values of 0.05, 0.01, respectively with n = 3. qRT-PCR analysis showed elevated gene expression of key gluconeogenic enzymes such as Pck2 (G) and Fbp2 (H) after transfecting EC4 cells with miR-122 mimic (miR-122) or NC mimic (NC). * and ** denote P values of ≤0.05 and 0.01, respectively (n = 3).