Fig. 4

Post-transcriptional regulation of Slc17a6. a Representative western blot of VGLUT2 (65 kDa) in the adult male hippocampus (n = 2 per group). GAPDH (38 kDa) was used as a loading control. b Quantitation of VGLUT2 levels in controls (n = 10 mice), basal expressors of Slc17a6 in the ethanol-exposed group (n = 4 mice) and high expressors of Slc17a6 in the ethanol-exposed group (n = 6 mice). c qPCR analysis of miRNA expression in the adult male hippocampus (n = 28 in the ethanol-exposed group and n = 32 in the controls). Expression was normalised to U6 snRNA. d miR-467b-5p targets the 3′UTR of Slc17a6 in an in vitro reporter assay. Luminescence is shown for vector only, vector containing the 3′UTR of Slc17a6 (Target) and vector with a mutated Slc17a6 3′UTR (Scramble) under three different co-transfection conditions: no co-transfection (None), addition of a miR-467b-5p mimic (Mimic) or addition of a miR-467b-5p inhibitor (Inhibitor). Assays were done in quadruplicate, and the results from three independent experiments in CAD cells are shown. Data are shown as mean and standard deviation. *P < 0.05, **P < 0.01 (t test, two-tailed)