Figure 6
From: The orphan nuclear receptor NR4A2 is part of a p53–microRNA-34 network
Overexpression of NR4A2 attenuates Nutlin-3a sensitivity.
HCT116 cells were transduced for 16 h with lentivirus expressing empty vector (EV) or 3xFlag-NR4A2 (NR4A2). The cell medium was then changed and the cells remained in culture for a total of 48 h. The cells were then reseeded into plates and the cell confluence was monitored using an IncuCyte ZOOM imaging system. After 14 h, the cell medium was replaced by medium containing vehicle (DMSO) or Nutlin-3a (5 or 10 μM) (indicated by Tx and arrow). The cell confluence for EV-expressing (a) or NR4A2-expressing (b) cells was monitored for a total of 108 h. (c) The cells were then collected and expression of NR4A2 was determined using TaqMan qPCR probes (normalized to ACTB). The value for DMSO-treated EV was set as 1. (d) Total protein lysates (35 μg) were assessed for expression of NR4A2 (low and high exposure), Flag (to indicate overexpressed NR4A2) and β-actin.
