Data for: Plasmid BMP-2–Embedded Gelatin Sponge As A Gene-Activated Matrix for Preosteoblast Differentiation
Description
Supplementary Figure 1 To allow endocytosis of DNA complexes into the cells, 5 × 104 cells and TransIT-2020/pEGFP-C1 complexes were mixed for 5, 15, 30, or 60 min (n = 3). Flow cytometry analysis was performed to evaluate GFP expression at these time points using FACS Calibur flow cytometer (Becton Dickinson, Heidelberg, Germany). Cells were gated at 1 × 104 cells/sample in FL1 channel setting, and BMP-2 transfection samples under the same condition at each time point were used as controls(Figure S1A). Transfection efficiencies of these four mixing time points were compared using Becton Dickinson Cell Quest software(Figure S1B). We found that duration of 30 min was the optimal mixing time, as the average GFP expression (9.92%) at 30 min was the highest compared to that at the other three time points. Live and dead assay : The scaffolds were stained using a Live/Dead Cytotoxicity/Viability kit (Invitrogen, Carlsbad, CA)from day 7 to day 28 .The live cells ,dead cells and merge images were all included. Figure 5 ALP supplementary data : all ALP activity corresponding to Figure 5 Figure 6 ARS supplementary data : all ARS activity corresponding to Figure 6 Figure 7 BMP-2 supplementary data: all BMP-2 expression corresponding to Figure 7