GTIN regulated Akt/NF-κB signaling pathways in VSMCs. A7r5 cells were incubated with GTIN at the indicated doses (0, 1, 5 and 10 μM) for 48 h. (a) The cytoplasmic levels of p-Akt, Akt, p-ERK, ERK and β-actin, as a cytosol internal control, were assayed by WB. (b) The nuclear levels of NF-κB, c-Jun, c-Fos and C23, as a nuclear internal control, were assayed by WB. The quantitative data was presented as means ± SD of three repeats from three independent studies. * p < 0.05, ** p < 0.01 compared with the untreated control via one-way ANOVA with Dunnett’s post-hoc analysis. (c) The DNA-binding abilities of nuclear NF-κB (c) and AP-1 (d) were analyzed using EMSA method. For the binding specificity controlling, Lane 1 represented the nuclear extracts reactive to free probe (the so-called unlabeled oligonucleotide). The results of EMSA were showed at least three independent experiments.