Fig. 2
Construction of lentiviral expression vectors and the establishment of a cell line of DA-BMSCs. (A) Dopamine biosynthetic processes involvement in DA-ergic presynaptic and postsynaptic neurons. Dopamine receptor subtypes (D1–D5); DAT dopamine transporter. (B) Schematic diagram of lentiviral vectors of LV-TH and LV-DDC-GCH1. (C) Fluorescent protein expression of BMSCs 72 h after lentivirus infection of LV-TH and LV-DDC-GCH1. LV-mCherry and LV-ZsGreen represented the control lentivirus. (D) The infection efficiency of the above lentivirus (n = 6). (E) Morphological characteristics of DA-BMSCs (P1–P4) and the expression of fluorescent proteins after puromycin and Blasticidin S selection. (F) Western blotting of TH, DDC, and GCH1 proteins in different lentivirus-infected cells. (G) RT-PCR analysis of TH, DDC, and GCH1 genes in different lentivirus-infected cells. (H) The relative mRNA expression of TH, DDC, and GCH1 was analyzed by RT-qPCR. (I) DA concentrations in the cells culture supernatant of each group were detected by HPLC. (J) HPLC chromatogram of cells culture supernatant was presented. Scale bars represent 100 μm. All data were presented as mean ± SEM. The P-values are from multiple comparisons in one-way ANOVA with Dunnett’s multiple comparison test (n = 3) ***P < 0.001, ****P < 0.0001. The original blots and gels are presented in Supplementary Fig. 2 (Fig. S2).
