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. 2021 Apr 20;9(1):e002104. doi: 10.1136/bmjdrc-2020-002104

Figure 5.

Figure 5

ICV of tofogliflozin altered c-fos expression and AMPK phosphorylation. At 1.5 hours after ICV infusion of 0.52 µmol/3 µL of tofogliflozin or vehicle into the LV, tissue samples were collected. (A) Expression of c-fos in the ARC and the PVH was assessed by RT-PCR and normalized to the expression of 18 s ribosomal RNA. (B) c-fos and β-tubulin expression were evaluated by immunoblotting. The results are shown as representative bands. All bands were quantified by densitometry. Band intensities of c-fos were normalized to β-tubulin. (C) Expression levels of total AMPK and AMPK phosphorylated at Thr172 were evaluated by immunoblotting. Results are shown as representative bands, and band are quantified by densitometry. Band intensities of phospho-AMPK were normalized to those of AMPK. (D) Expression of NPY, AgRP, and POMC in the ARC was assessed by RT-PCR and normalized to the expression of 18s ribosomal RNA. Data are presented as the mean±SD. Significance relative to vehicle control: *p<0.05, **p<0.01. V: vehicle, 2-DG: 2-deoxy-D-glucose, T: tofogliflozin A: (T, 2-DG, (V) n=4, 5, and 4, respectively. B and C: (V, T): n=3 and 5, respectively. D: (V, T) n=4 and 4, respectively. AgRP, agouti-related protein; ARC, arcuate nucleus; ICV, intracerebroventricular administration; LH, lateral hypothalamus; LV, lateral ventricle; NPY, neuropeptide Y; POMC, pro-opiomelanocortin; PVH, paraventricular nucleus; VMH, ventromedial hypothalamic.