Fig. 1

Schematic of DNA Vectors, Conditions, and Experimental Design for this Study.  a  Representative schematics of DNA vectors used in this study along with approximate size of DNA vectors in kilobase pairs (kbp). Sequences for each DNA vector are available at https://www.addgene.org/Angela_Pannier/. White element: CMV promoter; red element: tdTomato; green element: EGFP; grey element: SV40 polyA signal; pink element: P2A-T2A; blue element: IRES. b  Conditions tested for expression of multiple transgenes in hMSCs. (i) Separate complex conditions consisted of forming complexes separately for each single transgene DNA vector (pEGFP or ptdTomato, denoted as [pE]+[pT]). (ii) Same complex conditions consisted of forming complexes with both single transgene DNA vectors together (pEGFP + ptdTomato, denoted as [pE + pT]). Bi-cistronic (iii) D2A and (iv) IRES DNA vectors were formed in individual complexes. All conditions had equal mass of DNA as well as copy number of transgenes when directly compared amongst each other. Mass and copy number were equalized by addition of a promoterless pEGFP plasmid where needed. c  hMSCs from four donors (D1, D2, D3, & D4) and two tissue sources (adipose and bone marrow; hAMSCs and hBMSCs, respectively) were transfected with the conditions shown in « b » 24 h after seeding (4,500 and 6,000 cells/well, respectively) and imaged for transfection efficiency of each transgene 24 h after transfection