Fig. 2
Transduction of rBMSC and gene and protein expression of SP and αCGRP. A) Murine cDNA for SP, αCGRP and lacZ was cloned into lentiviral expression vectors, and viral particle were generated using a 293FT producer cell line. Subsequently, naive rBMSC were transduced with these viruses to produce lacZ-, αCGRP- or SP (over-) expressing rBMSCs clones. B) Gene expression analysis for SP and αCGRP was performed to verify expression level in transduced rBMSC in comparison to lacZ control rBMSC. C, D) SP and αCGRP protein concentration was determined in the cell culture supernatant of transduced rBMSC and adjusted to the secreted level from rBMSC-lacZ. E, F) SP and αCGRP protein concentration was determined in 3.5 × 105 transduced rBMSC, which corresponds to the number of cells per i.a. injection. Protein concentration of the neuropeptides was compared to that of the rBMSC-lacZ group. Results are means +/- SD; one sample t-test **p ≤ 0,01; N = 7–9