Journal: Cell Discovery
Article Title: DJ-1 maintains energy and glucose homeostasis by regulating the function of brown adipose tissue
Figure Lengend Snippet: Ablation of Akt1 mitigates the obesity and BAT dysfunction induced by DJ-1 transgene. ( a , b ) The effect of BYL-719 on gene and protein expression in BAT. ( c ) HE or anti-Ucp1 antibody staining in BAT was performed after BYL-719 treatment. ( d ) Body mass of wild-type mice (WT), DJ-1 transgenic mice (Tg, Line 17 C57BL/6J background), Akt1 KO mice (Akt1 −/− ), DJ-1 transgenic and Akt1 knockout mice (Akt1 −/− ; Tg) fed on a chow diet for 24 weeks (WT, n =15; Akt1 −/− , n =9; Tg, n =14; Akt1 −/− ; Tg, n =17). ( e ) Body lean and fat mass of 12-month-old indicated genotype mice measured by MRI (WT, n =7; Akt1 −/− , n =11; Tg, n =8; Akt1 −/− ; Tg, n =8). ( f ) Statistical analysis by General Linear Model (GLM) and ANCOVA analysis of the indicated genotype mice energy expenditure (EE) (WT, n =9; Akt1 −/− , n =8; Akt1 −/− ; Tg, n =9). ( g ) Lysates of BAT from mice of the genotypes indicated immunoblotted with Ucp1, Akt1, DJ-1 or β-tubulin antibodies. ( h ) HE and Ucp1 staining of BAT from 4-week-old mice of the indicated genotypes. ( i ) GTT results for 9-month-old indicated genotype mice fed on a chow diet (WT, n =8; Tg, n =7; Akt1 −/− , n =7; Akt1 −/− ; Tg, n =8).
Article Snippet: The antibodies used were anti-phosphor-Akt (Ser473; #4060, Cell Signaling, Cambridge, MA, USA), anti-Akt (#4691, Cell Signaling), anti-Akt1 (#2938, Cell Signaling), anti-FoxO1 (sc-11350, Santa Cruz, Dallas, TX, USA), anti-Akt2 (#3063, Cell Signaling), anti-phosphor-FoxO (Thr24-FoxO1/ Thr32-FoxO3a; #9464, Cell Signaling), anti-phosphor-HSL (Ser660; #4126, Cell Signaling), anti-HSL (#4107, Cell Signaling), anti-PTEN (#9188, Cell Signaling), anti-DJ-1 (#5933, Cell Signaling), anti-Ucp1 (ab10983), anti-Ubiquitin (#3936, Cell Signaling), anti-Mib2 (Ls-C295379, LifeSpan BioSciences, Seattle, WA, USA), anti-FLAG (Sigma), anti-GFP (Invitrogen), anti-HA (Santa Cruz), anti-β-tubulin (CW0098A, CWBiotech, Beijing, China), anti-β-actin (60008-1-Ig, Proteintech Group, Campbell Park, Chicago, IL, USA) and anti-GAPDH (CW0266A, CWBiotech).
Techniques: Expressing, Staining, Mouse Assay, Transgenic Assay, Knock-Out, Magnetic Resonance Imaging