The compound 7, 8-DHF was purchased from Sigma (⩾98%). 7, 8-DHF was dissolved in PBS with 17% dimethyl sulfoxide (DMSO). BMSCs were seeded into 6-well culture plates at a density of (2 × 107 cells/well) and cultured to ~90% confluence. BMSCs were treated with 7, 8-DHF using an optimal concentration of 500 nMol/L, as determined in a previous study, for 72 h.24 (link)
Fresh culture medium was replaced, and the BMSCs were separated into four experimental groups: (1) a negative control (NC) group, treated with medium only, (2) cells treated with 7, 8-DHF, (3) 7, 8-DHF + U0126 group, the cells were treated with 7, 8-DHF and U0126 (selective inhibitor of ERK1/2 signaling, 5 μM, Sigma-Aldrich, St. Louis, MO, USA), and (4) BDNF group, the cells were treated with human recombinant BDNF (159 nMol/L, Sigma).18 (link)
Following the treatment period, protein was isolated from the BMSCs for ELISA and Western blot analysis.
Fresh culture medium was replaced, and the BMSCs were separated into four experimental groups: (1) a negative control (NC) group, treated with medium only, (2) cells treated with 7, 8-DHF, (3) 7, 8-DHF + U0126 group, the cells were treated with 7, 8-DHF and U0126 (selective inhibitor of ERK1/2 signaling, 5 μM, Sigma-Aldrich, St. Louis, MO, USA), and (4) BDNF group, the cells were treated with human recombinant BDNF (159 nMol/L, Sigma).18 (link)
Following the treatment period, protein was isolated from the BMSCs for ELISA and Western blot analysis.
