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. 2020 Oct 28;10:570733. doi: 10.3389/fonc.2020.570733

Figure 7.

Figure 7

miR-550a-5p negatively regulated LIMD1 by directly binding to its 3′-UTR. (A) LIMD1 mRNA in 29 LUAD tissues and paired adjacent tissues was investigated by RT-qPCR. (B) A negative correlation was found between LIMD1 and miR-550a-5p in 29 clinical tumor samples. (C–E) Regulatory effect of miR-550a-5p on LIMD1 was detected by Western blot and RT-qPCR; GAPDH was used as an internal control for Western blot, while U6 was used for RT-qPCR. (F) The potential miR-550a-5p seed region at the 3′-UTR of LIMD1 mRNA. (G) A luciferase reporter assay was conducted to verify that miR-550a-5p directly bound to the 3′-UTR sequences of LIMD1. 1: pGL3-LIMD1; 2: pGL3-LIMD1 + miR-550a-5p mimics; 3: pGL3-LIMD1 + NC; 4: pGL3-LIMD1 mut; 5: pGL3-LIMD1 mut + miR-550a-5p mimics; 6: pGL3-LIMD1 mut + NC. Luciferase activity was normalized by the ratio of firefly and Renilla luciferase signals. Data expressed as mean ± SD (*P < 0.05; **P < 0.01; ***P < 0.001).