Figures & data
Figure 1. (A) Expression of ROBO1 in pancreatic cell lines. The relative quantitations of ROBO1 in BxPC-3-LN, BxPC-3, Panc-1, and SW1990 were 1.129 ± 0.1216, 0.3060 ± 0.8528, 0.3020 ± 0.06010, and 0.09967 ± 0.02255 respectively. (B) Expression of miRNA-218 in cells transfected with miRNA-218 precursor mimics. The relative quantitations of miRNA-218 in Blank Control group, Mimics-NC group, and Mimics-218 group were 1.311 ± 0.3077, 1.040 ± 0.3542, and 146.0 ± 13.48 respectively. (C) Expression of ROBO1 in cells transfected with miRNA-218 precursor mimics. The relative quantitation of ROBO1 in three groups were 1.007 ± 0.1475, 0.9970 ± 0.2872, and 0.2445 ± 0.03112 respectively. (D) The expression of ROBO1 protein in cells transfected with miRNA-218 precursor mimics. A significant decrease of ROBO1 protein was showed in Mimics-218 group. * represented the P value < 0.05.
Figure 2. (A) The predicted binding sites of miRNA-218 in the 3′UTR region of ROBO1. (B) miRNA-218 precursor mimics and pLuc-ROBO1-wt/mu were co-transfected into cells. The relative luciferase activities were 3.205 ± 0.2193 and 0.4857 ± 0.1466 in cells transfected with Mimics-NC+pLuc-ROBO1-wt, and Mimics-218+pLuc-ROBO1-wt respectively. The relative luciferase activities were 3.331 ± 0.2275 and 3.151 ± 0.2972 in cells transfected with Mimics-NC+pLuc-ROBO1-mu, and Mimics-218+pLuc-ROBO1-mu respectively. * represented the P value < 0.05.
Figure 3. (A–D) Cells transfected with Lenti-218 (A andC) or Lenti-NC (B andD) in regular optical vision and GFP vision (original magnification 100×). (E) Expression of miRNA-218 in cells transfected with lenti-218 or control. The relative quantitations of miRNA-218 in BxPC-3-LN group (blank control), Lenti-NC group, and Lenti-218 group were 1.007 ± 0.1466, 0.7627 ± 0.1704, and 143.5 ± 9.142 respectively. (F) The expression of ROBO1 protein in cells transfected with lenti-218 or control. A lower expressing level of ROBO1 protein was showed in Lenti-218 group compared with Lenti-NC group or Blank control. (G) Expression of ROBO1 in cells transfected with Lenti-218 or control. The relative quantitations of ROBO1 in three groups were 1.024 ± 0.2723, 0.9920 ± 0.1863, and 0.1327 ± 0.04005 respectively. P value < 0.05 was denoted as *.
Figure 4. (A–D) Cells transfected with Lenti-218 (A) or Lenti-NC (B), migrated to the inferior surface of the transwell inserts in GFP vision (original magnification 100×). Cells transfected with Lenti-218 (C) or Lenti-NC (D), invaded to the inferior surface of the transwell inserts in GFP vision (original magnification 100×). (E) The mean migrated or invaded cell counts per field in each group. The mean migrated cell counts in Lenti-218 and Lenti-NC group were 7.100 ± 2.601 and 22.20 ± 3.584 respectively. The mean invaded cell counts in Lenti-218 and Lenti-NC group were 1.800 ± 1.317 and 9.000 ± 2.211 respectively. (F) The expression of ROBO1 protein in cells transfected with pcDNA-muROBO1 or control. An increased expressing level of ROBO1 protein was showed in pcDNA-muROBO1 group compared with pcDNA-NC group. (G) Expression of ROBO1 in cells transfected with pcDNA-muROBO1 or control. The relative quantitations of ROBO1 in pcDNA-muROBO1 group and pcDNA-NC group were 1.913 ± 0.1149 and 0.1307 ± 0.03800 respectively. (H–K) Cells transfected with pcDNA-NC (H) or pcDNA-muROBO1 (I), migrated to the inferior surface of the transwell inserts in GFP vision (original magnification 100×). Cells transfected with pcDNA-NC (J) or pcDNA-muROBO1 (K), invaded to the inferior surface of the transwell inserts in GFP vision (original magnification 100×). (L) The mean migrated or invaded cell counts per field in each group. The mean migrated cell counts in pcDNA-NC and pcDNA-muROBO1 group were 11.70 ± 3.917 and 36.30 ± 4.832 respectively. The mean invaded cell counts in pcDNA-NC and pcDNA-muROBO1 group were 5.300 ± 2.214 and 21.80 ± 3.259 respectively. P value < 0.05 was denoted as *.
