Figure 5
From: The orphan nuclear receptor NR4A2 is part of a p53–microRNA-34 network
Knockdown of NR4A2 enhances p53 activation.
HCT116 cells were transfected for 24 h with nontargeting control (NT) or siRNA targeting NR4A2 (siNR4A2). The culture medium was then changed and the cells remained in culture for a total of 48 h. The cells were then treated with vehicle (DMSO) or Nutlin-3a (10 μM) for 24 h. Expression of NR4A2 (a) CDKN1A (b) MDM2 (c) mir-34a (d) and TP53 (e) was determined using TaqMan qPCR probes (normalized to GAPDH). The value for DMSO-treated NT was set as 1. The statistical significance of the results was determined using a two-way ANOVA with Tukey’s multiple comparison test. The significance of the differences within each transfection group between transfection groups (####P ≤ 0.0001; ###P ≤ 0.001; ##P ≤ 0.01) is represented. (f) HCT116 cells were transfected with NT or siNR4A2 as indicated above, followed by treatment of DMSO or Nutlin-3a (10 μM) for indicated length of times. Whole-cell lysates (45 μg) were assessed for expression of indicated proteins by performing Western blotting analysis.
