Figure 3.

HDACI inhibit glucose uptake and reduce GLUT1 expression before induction of apoptosis. A, Apoptosis (left) and glucose uptake (right) were analyzed in OPM2 cells treated with VPA, SAHA, or Dox. Apoptosis was analyzed as in Fig. 2A. For glucose uptake, 2 × 10⁵ live cells (per trypan blue exclusion) were incubated 10 min with [³H]2-DOG before washing and lysis of the cells. Retained radioactivity was detected by analysis in scintillation fluid. B, Apoptosis (left) and glucose uptake (right) in similarly treated H929 cells were analyzed as described in A. C, RNA isolated from OPM2 cells treated with 2 mm VPA was reverse transcribed before analysis by RTqPCR; detected levels of GLUT1 mRNA were normalized to the similarly detected housekeeping gene 36B4 mRNA. Fold induction over control was determined by setting GLUT1 levels in untreated cells equal to 1. D, GLUT1 mRNA levels in OPM2 cells treated 24 h with VPA, SAHA, or Dox (100 nm) was analyzed as in C. E, Lysates from OPM2 cells treated with 1 mm VPA were analyzed by immunoblot for expression of GLUT1 and loading control GAPDH. Results are representative of at least three independent experiments, and values represent calculated mean ± sd of triplicate samples.