FIG. 1.
Structure of the pH1RNAi vector and efficiency of gene silencing. (A) Structure of the pH1RNAi vector. The 19-mer siRNA target sequence is shown as N19. The sequence of the linker (ttcaagaga) and five thymidines (t) signaling termination are also shown. The transcribed RNA molecules self anneal to form hairpin-shaped short interfering RNA. (B) Silencing effect of the pH1Lama1 vector on laminin α1 expression at the mRNA level. The expression level of laminin α1 mRNA estimated by qPCR in cells transfected with the pH1Lama1 vector (pH1Lama1) is shown as a percentage relative to that in control cells transfected with the pH1S vector (control). The means ± SD of results from triplicate transfections are shown. (C) Silencing effect of the pH1Lama1 vector on laminin α1 expression at the protein level. Western blotting for laminin-1 proteins in conditioned medium from control cells or cells transfected with pH1Lama1 is shown. The solid arrowhead indicates laminin-1 (heterotrimer of α1, β1, and γ1 subunits), and the open arrowhead indicates either the α1 monomer or a heterodimer of β1 and γ1.