Figure 9.

Enhanced neuronal glycolysis via Pfkfb3 overexpression results in elevated MG levels. Pfkfb3 was overexpressed in neurons using lentiviral vectors and the following measurements were made 1 week later. A, Top panel, Pfkfb3 overexpression was confirmed by Western blotting; bottom panel, glucose utilization as measured by [³H]-2-DG uptake (data are expressed as a percentage of control values and are means ± SEM of at least 9 determinations from at least 3 independent experiments; control values were 211.8 ± 29.5 fmol/mg protein/20 min). B, Lactate release into the extracellular medium (data are expressed as percentage of control values and are means ± SEM of at least 9 determinations from at least 3 independent experiments; control values were 282.9 ± 50.1 nmol/mg protein/20 min). C, MG levels by HPLC (values are means ± SEM of at least 10 determinations from at least 4 independent experiments). A–C, Data were statistically analyzed using a Student's t test (**p ≤ 0.01; ***p ≤ 0.001). D, Western blot analysis shows increased neuronal argpyrimidine levels following Pfkfb3 overexpression. Representative bands are shown. Similar results were obtained in three independent experiments (n ≥ 8).