Figure 3

Over-expression of AAH and not Humbug increases motility in SH-Sy5y cells. SH-Sy5y cells were transfected with recombinant plasmid DNA to express luciferase (pLuc), AAH (pAAH), or Humbug (pHMBG). 48 hours after transfection during the peak period of gene expression, the cells were harvested to measure (A) AAH and (B) Humbug expression by real time quantitative RT-PCR with levels were normalized to 18S, and (C) directional motility using the ATPLite luminescence-based motility/invasion (ALMI) assay (see Methods). For the ALMI assays, percent motile refers to the combined mean percentage of motile adherent+motile non-adherent populations. The graphs depict the mean ± S.D. of results obtained from 6 or 8 replicate independent cultures. Within group comparisons were made using ANOVA with post-hoc Fisher Least Significant Difference (LSD) tests (* P < 0.001 relative to the other groups studied).