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. 2015 Jan 16;6(6):3904–3917. doi: 10.18632/oncotarget.2880

Figure 4. PAK6 is a direct target of miR-23a.

Figure 4

(A) Schematic of predicted miR-23a binding sequence in PAK6 3′-UTR. PAK6 3′-UTR was mutated in complementary site for seed region of miR-23a as indicated. A human PAK6 3′-UTR fragment containing wild-type or mutant miR-23a binding sequence was cloned downstream of luciferase reporter gene. (B) Real-time PCR was performed to detect PAK6 expression in PC-3 cells infected with miR-23a-expression vector or with empty vector. Data were normalized to GAPDH mRNA expression. (C) Luciferase activity of wild-type (Wt) or mutant (Mut) PAK6 3′UTR reporter gene in PC-3 cells infected with miR-23a-expression vector or empty vector. (D) PAK6 immunoblotting in PC-3 cells infected with miR-23a or with control RNA duplex (miR-control) and with anti-miR-23a (miR-23a inhibitor) or with anti-miR-control RNA duplex (negative control). (E) miR-23a expression was inversely correlated with PAK6 protein expression in 20 paired prostate cancer and adjacent non-tumor tissues. (F) Effects on invasion and migration of PC-3, DU145, C4-2 and C4-2B cells were rescued by overexpressing PAK6, which was consistent with immunoblotting results.