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. 2019 Dec 9;27(6):1844–1861. doi: 10.1038/s41418-019-0467-6

Fig. 6. Linc-ASEN binding to p21 mRNA through complementary base pairing is critical for p21 mRNA degradation.

Fig. 6

a Schematic representation of the pcFluc-p21 3′UTR (Wt) or pcFluc-p21 3′UTR ΔLinc-ASEN BS (Mut) (upper). MCF7 cells were transfected with pcFluc-p21 3′UTR Wt or Mut in combination with pRL-CMV as a reference plasmid, and then RT-qPCR analysis was performed (lower). b Diagrams of pcFLuc-MS2BS-p21 3′UTR (Wt) or pcFLuc-MS2BS-p21 3′UTR ΔLinc-ASEN BS (Mut) (upper). Using the MS2 tethering system, IP was performed, and precipitates were analyzed by western blot and RT-qPCR (lower). c Schematic representations of the pCMV-Linc-ASEN (Wt) or pCMV-Linc-ASEN Δp21 RBS (Mut) (upper). Relative Linc-ASEN and p21 mRNA levels were analyzed by RT-qPCR (lower). d MCF7 cells were transfected with pCMV-Linc-ASEN-MS2BS (Wt) or pCMV-Linc-ASEN Δp21 RBS-MS2BS (Mut) reporter plasmids in combination with pMS2-HA. Cells were subjected to IP and western blot analyses. Precipitated RNA was subjected to RT-qPCR. In a–d, data are represented as means ± SD. *P < 0.05; **P < 0.01; #P > 0.05 (Student’s t-test).