Fig. 3

SV40 large T antigen, DTT and 2-DG promote alternative TrkAIII splicing in NB cells. a) Representative RT-PCR agarose gel demonstrating the promotion of sequence verified alternative TrkAIII expression in SH-SY5Y NB cells, following transient transfection with plasmids pw2dl expressing SV40 large T-ag alone (T+/t-), pw2t expressing SV40 small tag alone (T−/t+), pw2 expressing both large T-ag and small t-ag (T+/t+) and negative control pw101 expressing only mutation inactivated small t-ag (T−/t-) compared to non-transfected control (Con). Large T-ag expression was confirmed by RT PCR amplification using T-ag specific primers whilst small tag expression was confirmed by deduction using a primer set that generates an identical fragment form both large T-ag and small tag cDNAs. b) Representative RT-PCR agarose gels demonstrating the promotion of alternative TrkAIII splicing in SH-SY5Y cells treated with 1 mM DTT for 3–12 h plus DTT-induced Xbp-1 splicing, CHOP and Grp78/Bip expression compared to GAPDH levels. c) Representative RT-PCR demonstrating induction of alternative TrkAIII splicing in SH-SY5Y cells treated for 6 and 12 h with the glucose deprivation mimic 2 deoxy-glucose (1 mM 2-DG) plus TrkAIII cDNA and GAPDH RT PCR controls