RT–PCR analysis for GDNF , NGF and VEGF (A, B and C, respectively). ATB-346 treatment significantly increased both GDNF and NGF mRNA expression compared to TBI (A and B, respectively). ATB-346 determined an important increase in VEGF mRNA expression (C). β-actin was used as an internal control. mRNA was extracted and reverse-transcribed as described in the Methods section. Similar results were obtained in four additional separate experiments. No bands were observed in the absence of cDNA. Western blot analysis showed that eNOS expression in TBI mice was increased compared with sham mice (D), while ATB-346 upregulated its expression (D). A representative blot of homogenates obtained from five animals per group is shown, and densitometry analysis of all animals is reported (A1 to D1). A P value of less than 0.05 was considered significant. *P < 0.05, **P <0.01, ***P <0.001 versus sham, #P <0.05 versus TBI.