Figure 1.

Characterization of cultured rat BMMSCs and detection of lentivirus-mediated overexpression of BDNF and VEGF. (a) Representative phase-contrast micrographs show the morphology of rat BMMSC cultures at passage 0 (P0) and passage 3 (P3); (b) Flow cytometry analysis shows the majority of BMMSCs at P3-expressing stem cells markers CD44 and CD90, and a small portion of cells expressing CD34 and CD44; (c) Schematics (on the top) show the expression cassette with either rat brain BDNF exon IV or VEGF-A open reading frame coding sequence followed by an IRES-directed ZsGreen1 and tdTomato fluorescent protein coding sequences in the lentivirus constructs, respectively; fluorescence micrographs (four panels in the bottom) show that transduced BMMSCs co-express both ZsGreen1 (green) and tdTomato (red) fluorescent proteins; (d) RT-qPCR results show the relative abundance of BDNF and VEGF mRNAs in un-transduced (naive) BMMSCs as well as 48 h after transduction with an empty lentiviral vector (Lent-vector) or BDNF-&VEGF lentivirus-co-transduced (Lenti-B&V). Data are depicted as mean±S.D.; (e) Western blots of whole-cell lysates detect markedly increased levels of BDNF and VEGF in cells co-transduced with both BDNF- and VEGF-lentiviruses