Figure 2

NGF is a key to mediate BMSC-afforded neuroprotection. (A) The protein levels of NGF were detected in the spinal cord and sciatic nerve of rats by using commercial ELISA kit. (B) Neutralization of NGF by using anti-NGF antibody abolishes the inhibitory effects of BMSC against HD-induced elevation of LC3II. VSC4.1 cells were treated with HD (25 mM) or saline for 24 h and then were treated with BMSC-CM (15%, v/v) or NGF (100 μM) in the presence or absence of anti-NGF antibody (10 μM) for additional 24 h. The levels of LC3 determined by western blot and the density of blots were quantified (the full-length gels were shown in Supplementary Figure S3A). (C) LC3 was stained in different groups and the representative images were shown. (D) TEM was used to detect the autophagosome in different groups. (E) K252a, the inhibitor of TrkA, attenuates the inhibitory effects of BMSC against HD-induced elevation of LC3II. VSC4.1 cells were treated with HD (25 mM) or saline for 24 h and then were treated with BMSC-CM (15%, v/v) with or without 1 h pre-treatment of the inhibitor of TrkA, K252a, for additional 24 h. The levels of LC3 determined by western blot and the density of blots were quantified (the full-length gels were shown in Supplementary Figure S3B). ^ap < 0.05, compared with control group; ^bp < 0.05, compared with HD group; ^cp < 0.05, compared with BMSC-CM group.