Figure 3. KDM1 inhibition enhanced p53 functions and its target gene activation.

(A) Whole cell lysates were isolated from vehicle-, pargyline- or NCL-1 treated U87 and LN229 cells and subjected to Western blot analysis with p53 and acetyl-p53³⁸² antibodies (upper panel). Band intensity of acetyl p53 was quantitated by densitometry and normalized to total p53. (B) U87 and LN229 cells were transiently transfected with the p53-Luc reporter and 24 h post transfection, cells were treated with vehicle, pargyline or NCL-1. The reporter gene activity was measured after 24 h. All data presented are the mean of 3 independent experiments with mean ± SEM. *, p< 0.05, t test. (C) U87 and LN229 cells were transfected with control siRNA or KDM1 siRNA and 72 h after transfection, RNA was isolated and subjected to RT-qPCR using the primers specific for p53 target genes p21 and PUMA. All data presented are the mean ± SEM. *, p< 0.05, t test. Total RNA was isolated from vehicle-, pargyline or NCL-1 treated U87 (D) and LN229 (E) cells and subjected to RT-qPCR using the primers specific for p21 and PUMA. All data presented are the mean ± SEM. *, p< 0.05, t test. (F) Whole cell lysates were isolated from vehicle-, pargyline- or NCL-1-treated U87 and LN229 cells and subjected Western blot analysis with p21 antibodies. β-actin used as an internal control. Band intensity of p21was quantitated by densitometry and normalized to total actin.