Fig. 5

Tas1r3 knockdown alleviates high glucose, fructose and palmitate-induced suppression of CREB/BDNF signaling in hypothalamic neuronal cell. A Study design of siRNA knockdown in hypothalamic neuronal cell line mHypoA-2/12. B Evaluation of siRNA-mediated Tas1r3 mRNA suppression in a cultured adult hypothalamic neuronal cell line after 48 h of transfection. Two-way ANOVA followed by Tukey’s multiple comparison test: ***p < 0.001. n = 4/group. C-E mHypoA-2/12 cells were transfected with Tas1r3 siRNA or scrambled control siRNA for 48 h and stimulated with or without fructose (10 mM), glucose (10 mM), and palmitate (10 μM) for 24 h. Relative mRNA expression of Prkaca, Creb1, and Bdnf. Two-way ANOVA followed by Tukey’s multiple comparison test: *p < 0.05, **p < 0.01, and ***p < 0.001. n = 3/group. F Protein levels of BDNF released into the supernatant. Two-way ANOVA followed by Tukey’s multiple comparison test: **p < 0.01, ***p < 0.001. n = 3/group. G mHypoA-2/12 cell viability at 0, 12, and 24 h. Two-way ANOVA followed by Tukey’s multiple comparison test: *p < 0.05, **p < 0.01. n = 3/group. H,I Relative mRNA expression of Map2 and Dcx. Two-way ANOVA followed by Tukey’s multiple comparison test: *p < 0.05, **p < 0.01, and ***p < 0.001. n = 3/group. All data are presented as mean ± SEM. ANOVA, analysis of variance; BDNF, brain-derived neurotrophic factor; CREB, cAMP response element-binding protein