Fig. 3

Delivery of Bi-Cistronic DNA Vectors with an IRES or D2A Sequence for Expression of Two Reporter Transgenes in hMSCs.  hMSCs were transfected with bi-cistronic DNA vectors complexed with Lipofectamine 3000 (a, c, e, & g) or Turbofect (b, d, f, & h) and transfection efficiencies for each transgene (EGFP, green bars, or tdTomato, red bars) were compared to the expression levels of single transgene vectors (pEGFP or ptdTomato) in D1 hAMSCs (a & b), D2 hAMSCs (c & d), D3 hBMSCs (e & f), and D4 hBMSCs (g & h). Transfection efficiencies for hMSCs expressing both EGFP and tdTomato (co-expression, yellow bars) was calculated by dividing the number of cells that were both EGFP and tdTomato positive by the total cell count (Hoechst, nuclear stain). All conditions have equal moles of expression cassette and mass of DNA delivered. 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