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. 2019 Jul 29;20(15):3705. doi: 10.3390/ijms20153705

Figure 3.

Figure 3

ESRP1 overexpression leads to decreased cyclin A2 mRNA stability. (A) 48 h after transfection with an ESRP1 vector, ESRP1 expression in HeLa cells was detected by qRT-PCR. (B) 24 h following ESRP1 vector transfection, cells were treated using 5 g/mL actinomycin D (AMD), with RNA being isolated at the indicated times, after which cyclin A2 expression was determined via qRT-PCR. (C) Schematic diagram of the construction of a luciferase-cyclin A2 3′UTR chimeric vector generated via subcloning the cyclin A2 3′UTR region into the pEZX-MT06 vector encoding firefly luciferase. (D) HeLa cells or HEK293 cells in 24-well plates were co-transfected with full-length vector or control vector, along with ESRP1 vector (0, 10, 100, and 1000 ng), and after 48 h, cells were lysed and luciferase activity was assessed. For normalization of luciferase activity, the signal (1 μg protein) for control cells was set to 1. This experiment was repeated three times (n = 3); error bars indicate standard error; ** p < 0.01.