Extended Data Fig. 3: Effect of systemic treatment with SF1-AMPKα1-DN loaded sEVs on hypothalamic AMPK activity in DIO mice.
a, Representative confocal images depicting GFAP (green), pACCα (magenta) and merged reactivity in brain sections (control n = 8 fields, 4 mice/group; SF1-AMPKα1-DN n = 8 fields, 4 mice/group) after 24 h of intravenous injection with control (non-loaded) or SF1-AMPKα1-DN loaded sEVs; scale bars represent 20 µm. b, Representative confocal images depicting Iba1 (green), pACCα (magenta) and merged reactivity in brain sections after 24 h of intravenous injection with control (non-loaded) or SF1-AMPKα1-DN loaded sEVs; scale bars represent 20 µm. c, Negative controls for pACCα and SF1 double immunofluorescence. Representative confocal images depicting DAPI (blue), Alexa594, with or without SF1 (red), Alexa 488 with or without pACCα (green) and merged reactivity in brain sections; scale bars represent 20 µm. The experiments were repeated 3 times. d, Quantification of pACCα fluorescence in ARC, DMH and PVH (quantification per field; ARC control n = 12 fields, 3 mice/group; SF1-AMPKα1-DN n = 8 fields, 2 mice/group; DMH control n = 12 fields, 3 mice/group; SF1-AMPKα1-DN n = 12 fields, 3 mice/group; PVH control n = 4 fields, 1 mice/group; SF1-AMPKα1-DN n = 12 fields, 3 mice/group) after 24 h of intravenous injection with control (non-loaded) or SF1-AMPKα1-DN loaded sEVs. Data expressed as mean ± SEM. Statistical significance was assessed by two-sided Student’s t-test.
