MTG16 inhibition of <i>PFKFB3, PFKFB4</i> and <i>PDK1</i> during hypoxia.

<p>The time course is shown for transcriptional expression of <i>MTG16</i>, <i>PFKFB3, PFKFB4</i>, and <i>PDK1</i> in Raji/MTG16 Tet-On 3G cells during incubation without doxycycline (open bars) or 20 ng/ml doxycycline (closed bars) under hypoxic conditions (4% O₂). Upregulation of <i>MTG16</i> expression by doxycycline was achieved also during hypoxia. Doxycycline induction of MTG16 diminished the hypoxia induction of <i>PFKFB3, PFKFB4</i> and <i>PDK1</i> expression. For protein detection, cell lysates were probed with antibodies to PFKFB3, PFKFB4, PDK1, β-actin and Histone H3 by Western blot analysis. The intensity of the bands was quantified by densitometry and expressed as ratios to β-actin or Histone H3. Results are shown after incubation with 20 ng/ml doxycycline for 0, 24 and 48 h during hypoxia. The Western blotting experiments were repeated at least three times and representative results are shown. The level of PFKFB4 and PDK1 was decreased by doxycycline–induction of MTG16. Data are represented as means±SEM for n = 3 and were compared by the two-way ANOVA followed by the Bonferroni post-hoc test (*p<0.05; ***p<0.001).