Figure 1

Generation of recombinant resistin lentivirus and detection of recombinant gene expression. (A) A full length of mouse resistin coding sequence (0.34 kb) was cloned into a ViraPower-CMV vector (Invitrogen). This recombinant resistin gene construct included a V5 epitope tag and a CMV promoter. (B) A western blot analysis was performed to confirm the expression of the recombinant resistin genes in three different stably transduced cell lines (R18, R20 and R23) using anti-V5 antibodies. The lanes marked by the letter C were loaded with proteins from non-transduced control 3T3-L1 cells. The lane marked by the letter Z was loaded with proteins from cells transduced lentivirus containing recombinant LacZ gene including the V-5 tag and LacZ was detected using anti-V5 antibodies. (C) A western blot analysis was performed to examine the secretion of the recombinant exogenous resistin proteins (R18, R20, and R23) and the endogenous resistin proteins in the medium of 3T3-L1 cells. Exogenous and endogenous resistin proteins were detected using anti-resistin antibodies. The control lane marked by the letter C was loaded with proteins from non-transduced 3T3-L1 cells.