Up-regulation of VEGF-B transcript, but not of VEGF-A transcript, after rotenone (20 nM) treatment of midbrain cultures. The fold-increase of transcript vs. control as determined with real time PCR is shown as mean ± s.e.m for VEGF-B (black bar) and VEGF-A (shaded bar); n = 7 separate experiments. Mean fold changes in rotenone-treated cultures compared to control (± s.e.m.) were: VEGF-B = 1.65 ± 0.29; P = 0.026; VEGF-A = 1.25 ± 0.20; P = 0.40. Statistical significance was determined by a two-tail t test of ΔΔCt and difference from control is depicted by an asterisk. Oligonucleotide Primers from Applied Biosystems Inc., Foster City, CA were used: VEGF-A (Rn01511602_m1); VEGF-B (Rn01454585_g1); β-actin (Rn00667869_m1). The TaqMan® MGB probe used at a concentration of 5 μM, comprised of a FAM™ reporter dye at the 5' end and a nonfluorescent quencher at the 3' end of the probe. The solution also contained the necessary primers at a concentration of 18 μM each.