Figure 5.

Increased FTO stabilizes the Coll IV microinjection-induced TLR4 upregulation in the thalamus. (A) Effect of thalamic pre-microinjection of AAV5-Cre or AAV5-Gfp into unilateral thalamus of Fto^fl/fl male mice 35 days before Coll IV or saline microinjection into the same regions on the expression of TLR4 protien in thalamus. n = 5 mice/group. Statistical significance was calculated with two-way ANOVA (treatment × model) followed by post hoc Tukey test. F_{treatment (1, 16)} = 65.941, p < 0.001; F_{model (1, 16)} = 60.536, p < 0.001; F_{treatment × treatment (1, 16)} = 52.71, p < 0.001. (B) Level of TLR4 protein in the ipsilateral thalamus 5 weeks after microinjection of AAV5-Fto or AAV5-Gfp into unilateral thalamus. n = 5 mice/treatment. Statistical significance was calculated with two-tailed unpaired Student’s t-test. t ₍₈₎ = −6.442. (C) Effect of intraperitoneal administration of meclofenamic acid (MA; 10 mg/kg) once daily for 5 days starting 35 days post-viral microinjection on the levels of FTO and TLR4 proteins in the ipsilateral thalamus of naïve male mice 40 days after microinjection. n = 5 mice/treatment. Statistical significance was calculated with two-way ANOVA (treatment × model) followed by post hoc Tukey test. FTO: F_{model (1, 16)} = 98.045, p < 0.001; F_{treatment (1, 16)} = 0.015, p = 0.904; F_{treatment × treatment (1, 16)} = 0.038, p = 0.848. TLR4: F_{model (1, 16)} = 38.893, p < 0.001; F_{treatment (1, 16)} = 27.386, p < 0.001; F_{treatment × model (1, 16)} = 27.313, p < 0.001. (D) Co-expression of Fto mRNA and Tlr4 mRNA in individual thalamic neurons. n = 3 mice. Tuba-1a was used a positive control.