Fig 2: miR-218 directly regulates LAMB3. (A) mRNA expression of LAMB3 as measured by qRT-PCR 72 h after transfection with 10 nM miR-218. GAPDH was used as an internal control. *P<0.0001. (B) Protein expression of LAMB3 as measured by western blot analysis 72 h after transfection with 10 nM miR-218. GAPDH was used as a loading control. The expression ratio of LAMB3 was evaluated using ImageJ software. (C) A putative miR-218 binding site in the 3′UTR of LAMB3 mRNA was identified using the TargetScan database (Upper). Luciferase reporter assays were performed using a vector encoding the partial sequences of the 3′UTR containing the putative miR-218 target site. The vector (10 ng) and miR-218 or miR-control (10 nM) were cotransfected into CaSki cells. Renila luciferase activity was measured 24 h after transfection. The results normalized by firefly luciferase values are shown. *P<0.0001.

Fig 3: Expression of LAMB3 in cervical-SCC clinical specimens. (A) Expression of LAMB3 in cervical-SCC tissues and nontumor tissues as determined by qRT-PCR. ACTB was used as an internal control. (B) The correlation between the expression of miR-218 and LAMB3 was analyzed in cervical-SCC and non-tumor tissues.

Fig 4: LAMB3 expression was suppressed by si-LAMB3 transfection at both the mRNA and protein levels in CaSKi cells. (A) mRNA expression of LAMB3 as revealed by real-time qRT-PCR 72 h after transfection with 10 nM si-LAMB3. *P<0.0001. (B) Protein expression of LAMB3 as revealed by western blot analysis 72 h after transfection with 10 nM si-LAMB3. GAPDH was used as a loading control. The expression ratio of LAMB3 was evaluated using ImageJ software.