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Serum was analyzed with spectrophotometric kits for NEFA, NEFA Fa-115 (Randox, London, UK), -hydroxybutyrate (-HB) by Ranbut RB 1008 (Randox, London, UK), and urea by UREA/BUN Color (Biosystems, Barcelona, Spain) concentrations

After washing with PBS, the cells were treated with Alexa Fluor 488-conjugated secondary antibodies (Invitrogen) for 1 h and subsequently stained with 4,6-diamidino-2-phenylindole (DAPI, Invitrogen) at room temperature

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