Figure 4
From: PTBP1-mediated regulation of AXL mRNA stability plays a role in lung tumorigenesis
Post-transcriptional control of AXL mRNA stability by PTBP1. (A) The transcription rate of AXL was similar in different cells. Nuclei from 2 × 106 cells were collected and incubated in a reaction buffer containing ATP, GTP, CTP and biotin-16-UTP and subjected to nonradioactive nuclear run-on assay. In some reactions (negative controls), UTP was used in place of biotin-16-UTP. Total RNA was isolated by TRIzol extraction, and biotinylated RNA was purified using agarose-conjugated streptavidin beads. Expression level of AXL mRNA was measured by quantitative real-time RT-PCR. Actin was used as the internal control. The mRNA stability measurement experiments were initiated by adding 10 μg/ml ActD to (B) Empty vector-transfected CL1-0 vs. PTBP1-transfected CL1-0 and to (C) Empty vector-transfected CL1-5 vs. PTBP1-transfected CL1-5, respectively. Cells were harvested at intervals as indicated and AXL mRNA decay was analyzed by quantitative real-time RT-PCR. The values were derived from three independent experiments. Error bars represent SD. *P < 0.05.
