Fig. 2.

Retained transduction and transcriptional activity of released P21- RUNX2-8R. PLGA MPs were prepared using S/O/W method, loaded with P21- RUNX2-8R-FITC (0.5% (w/w)). (A) Transduction activity of the released protein was assessed in hMSC monolayers. Representative fluorescence microscopy images of the hMSCs treated for 24 h with the released P21-RUNX2-8R-FITC (Scale bar represents 100 μm) collected on the days indicated. (B) Flow cytometry histograms showing cellular uptake of P21-RUNX2-8R-FITC in hMSCs. The green histograms are the released P21-RUNX2-8R-FITC, while the black histograms are fresh experimental controls of P21-RUNX2-8R-FITC delivered at the same released concentrations (between 3 and 20 μg/ml)(the same fresh controls were used for day 5 and 7). Gates of from control cells showing negative (left) and positive (right) cells. (C) Relative transduction activity of released P21- RUNX2-8R-FITC compared to fresh protein. Values are the mean fluorescence intensity (MFI) per cell from flow cytometry. (D) Transcriptional activity of the released P21-RUNX2-8R-FITC measured using mOG2-Luc RUNX2-responsive luciferase reporter gene assay. pRUNX2 (plasmid DNA, pSIN-RUNX2) transfection was used as a positive control. Values induced after 1 or 2 days transduction are presented. Error bars indicate SD (n = 3). ** p < .05. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)