Fig. 1: Production and characterization of engineered BMP2- and BMP7-EV. | npj Regenerative Medicine

Fig. 1: Production and characterization of engineered BMP2- and BMP7-EV.

From: Engineered BMP2/BMP7 extracellular vesicles induce autocrine BMP release driving SMAD phosphorylation to promote bone formation

Fig. 1: Production and characterization of engineered BMP2- and BMP7-EV.The alternative text for this image may have been generated using AI.

A Schematic of lentiviral vector with BMP2 (top) or BMP2 and BMP7 (bottom) joined with T2A linker. Acronyms are as follows: LTR (Long Terminal Repeat), Ψ (viral packaging), RRE (Rev response element), cPPT (central polypurine tract), WPRE (post-regulatory element), ΔU3 (U3 deletion in 3’ LTR). B Real-time quantitative reverse-transcriptase PCR to show ΔCT ratio to GAPDH in BMP2 or BMP7 mRNA in MSCs transfected with either LV-BMP2 or LV-BMP2/7 plasmid. Statistical significance determined by one-way ANOVA with Tukey post hoc analysis. (+/- SD, n = 3). C Western blot of MSCs that were untransfected or transfected with LV-BMP2 or LV-BMP2/7. Actin was used as a loading control. D NanoSight analysis demonstrating mean size profile (+/- SD) of extracellular vesicles isolated from LV-BMP2 or LV-BMP2/7 transfected MSCs. E Transmission electron microscopy (TEM) demonstrating size profile of BMP2-EV or BMP2/7-EV. Scale bar 200 nm. F Western blot of small extracellular vesicles collected from MSCs that were untransfected or transfected with LV-BMP2 or LV-BMP2/7. Samples were probed for small extracellular vesicle markers CD63, Flotillin, and CD9. G Western blot of small extracellular vesicles collected from MSCs that were untransfected or transfected with LV-BMP2 or LV-BMP2/7. Samples were probed for BMP2 and BMP7 with actin as a loading control.

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