Figure 4. Acute exercise improves S1PR1/STAT3 in the hypothalamus of rats. Experimental design (a). Single and double-immunostaining was performed to evaluate the co-localization of S1PR1 (green) with STAT3 phosphorylation and POMC (red) in the third ventricle young, Middle-Aged and Middle-Aged exercised of rats (b). The hypothalamic extraction for analysis was made 3 hours after the acute protocol exercise. Food intake over 12 hours (c) (n= 8 per group). Hypothalamic NPY mRNA levels (d) (n=6–7 per group). Western blot showing S1PR1 expression and STAT3 phosphorylation (e) in the hypothalamus of middle-aged vs. middle-aged exercised (n=6 per group). UCP1 mRNA levels in BAT (f) (n=6 per group). Western blot showing UCP1 expression in BAT (g) and Image of BAT (n=6 per group) (h). The samples of hypothalamus and BAT were extracted after three hours of exercise. The Student’s t-test was performed where (c) ***p<0.0003, (d) **p<0.0026 and (f) *p<0.0123.