Fig. 5

Equal Mass of DNA and Copy Number of Transgenes Comparison for Delivery of Two DNA Vectors to Delivery of Bi-Cistronic DNA Vectors for Expression of Two Transgenes in hMSCs.  hMSCs were transfected with single transgene DNA vectors delivered in separate complexes ([pE]+[pT]) the same complex ([pE + pT]) or bi-cistronic DNA vectors (pD2A or pIRES) with an equal mass of DNA and copy number of transgenes delivered. All four conditions were complexed with Lipofectamine 3000 (a, c, e, & f) or Turbofect (b, d, f, & h). Transgene expression for each condition was assayed 24 h after transfection for D1 hAMSCs (a & b), D2 hAMSCs (c & d), D3 hBMSCs (e & f), and D4 hBMSCs (g & h). Expression of both transgenes simultaneously in hMSCs (co-expression) was calculated by dividing the number of cells that were both EGFP and tdTomato positive by the total cell count (Hoechst, nuclear stain). Data represented as mean ± SEM (n = 6). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ns, not significant (p > 0.05), as determined by a 2-way ANOVA with Tukey’s post hoc test