Skip to main content
. 2015 Sep 25;6:8494. doi: 10.1038/ncomms9494

Figure 4. CXCR4 is a direct miR-622 target.

Figure 4

(a) Schema of the candidate miRNAs by different prediction algorithms. Each labelled circle represents one prediction algorithm with the number of its predicted miRNAs, and the number listed in overlapping of circles is simultaneously predicted by different algorithms. (b) Schematic model for miRNA screening to target CXCR4. (c) Heatmap obtained from RT–PCR of HCC and corresponding peritumour specimens. Each column represents the average of three biological replicates. The relative high expression is indicated in red, whereas the relative low expression is in green. (d) Western blot analysis of CXCR4 and GAPDH for SK-Hep1 cells transiently transfected with miR-nc, miR-302c, miR-139-5p, miR-9, miR-206 and miR-622 mimic (left panel) and for Huh7 cells transiently transfected with anti-miR-nc, anti-miR-302c, anti-miR-139-5p, anti-miR-9, anti-miR-206 and anti-miR-622 (right panel). Data are representative immunoblots of three independent assays. (e) Sequences of miR-622 and the potential miR-622-binding sites at the 3′-UTR of CXCR4. Also shown are nucleotides mutated in CXCR4-3′-UTR mutant. Seed sequences are marked. (f) Luciferase activity assay for pGL3-CXCR4 3′-UTR (wt) or pGL3-CXCR4 3′-UTR (mut) relative to Renilla luciferase activity for SK-Hep1 and SNU448 cells transiently transfected with miR-nc or miR-622-mimc (n=3, upper panel). Student's t-test, *P<0.05. Error bars in panels are defined as s.d. Western blot analysis of CXCR4 and GAPDH for SK-Hep1 and SNU448 cells transiently transfected with miR-nc and miR-622-mimc (lower panel). Data are representative immunoblots of three independent assays. (g) Luciferase activity assay for pGL3-CXCR4 3′-UTR (wt) or pGL3-CXCR4 3′-UTR (mut) relative to Renilla luciferase activity for PLC/PRF/5 and Huh7 cells transiently transfected with anti-miR-nc or anti-miR-622 (n=3, upper panel). Student's t-test, *P<0.05. Error bars in panels are defined as s.d. Western blot analysis of CXCR4 and GAPDH for PLC/PRF/5 and Huh7 cells transiently transfected with anti-miR-nc or anti-miR-622 (lower panel). Data are representative immunoblots of three independent assays.