injection with leptin  (R&D Systems)

Journal: Nature Communications

Article Title: Regulation of leptin signaling and diet-induced obesity by SEL1L-HRD1 ER-associated degradation in POMC expressing neurons

doi: 10.1038/s41467-024-52743-2

Figure Lengend Snippet: a Daily food intake of Sel1L f/f and Sel1L POMC male mice at 1w- and 8w-HFD (1w-HFD: n = 11, 9 mice for Sel1L f/f and Sel1L POMC ; 8w-HFD: n = 9, 11 mice for Sel1L f/f and Sel1L POMC ). b Growth of male Sel1L POMC mice under 8-week ad libitum, 8-week pair feeding and 1-week ad libitum on HFD ( n = 3 mice, blue dots). Male Sel1L f/f mice fed ad libitum with the same diets were included as controls ( n = 3 mice, black dots). c Growth of Sel1L f/f male mice ad libitum and Sel1L POMC male mice either ad libitum or pair-feeding of HFD starting from 5 weeks of age (ad libitum: n = 11, 8 mice for Sel1L f/f and Sel1L POMC ; pair feeding: n = 3 Sel1L POMC ). d , e , f Diagram of leptin sensitivity test ( d ). 12-week-old male mice on HFD were injected daily i.p. with vehicle PBS and then PBS or leptin (2 mg/kg body weight) for 3 days. e , f Body weight change ( e ), average daily food intake ( f ) following 3 daily vehicle or leptin injections of the male mice (PBS: n = 4 mice per group; leptin: n = 6, 4 mice for Sel1L f/f and Sel1L POMC ). Body weight change was calculated by end point body weights minus starting point body weights. g Serum leptin levels in male mice fed on NCD, 1w- and 8w-HFD (NCD: n = 13, 11 mice for Sel1L f/f and Sel1L POMC ; 1w-HFD: n = 7, 8 mice for Sel1L f/f and Sel1L POMC ; 8w-HFD: n = 5, 8 mice for Sel1L f/f and Sel1L POMC ). NCD for normal chow diet; HFD for high fat diet. Values, mean ± SEM. n.s. not significant; p values are as indicated unless **** p < 0.0001 by two-way ANOVA followed by Bonferroni’s multiple comparisons test ( a , b , e , f ) or Tukey’s multiple comparisons test ( c , g ). Source data are provided as a Source Data file.

Article Snippet: Twelve-week-old mice were acclimatized to single housing with HFD and intraperitoneal (i.p.) injection (PBS) for three days, followed by three daily leptin injections (2 mg/kg body weight, R&D systems; catalog 498-OB-05M) 1 h before the onset of dark cycle as described .

Techniques: Injection

Journal: Nature Communications

Article Title: Regulation of leptin signaling and diet-induced obesity by SEL1L-HRD1 ER-associated degradation in POMC expressing neurons

doi: 10.1038/s41467-024-52743-2

Figure Lengend Snippet: a – d Representative immunofluorescence (IF) staining of pSTAT3 (red) and eGFP (green) in Sel1L f/f ;Pomc-eGFP and Sel1L POMC ;Pomc-eGFP mice under NCD ( a ), 1w-HFD ( b ) and 8w-HFD ( c ). Mice were fasted overnight followed by i.p. injection with leptin (2 mg/kg body weight), with quantitation of percentage of pSTAT3 + POMC neurons shown in ( d ). Mice injected with PBS were included as negative controls (Supplemental Fig. ). Yellow arrows, pSTAT3 + POMC neurons; white arrowheads, pSTAT3 - POMC neurons. (NCD: n = 4 mice per genotype; 1w-, 8w-HFD and PBS: n = 3 mice per genotype). e Quantitative PCR (qPCR) analysis of Pomc mRNA levels in ARC of Sel1L f/f and Sel1L POMC male mice at 8w-HFD ( n = 3 mice per group). f , g Representative Western blot analysis ( f ) and quantitation ( g ) of leptin-induced pSTAT3 in ARC of Sel1L f/f and Sel1L POMC mice under NCD or 8w-HFD (NCD: n = 2 mice per genotype for PBS, n = 4 mice per genotype for leptin; 8w-HFD: n = 4 mice per group). NCD for normal chow diet; HFD for high fat diet; ARC, arcuate nucleus. arb. units, arbitrary units. Values, mean ± SEM. n.s. not significant; p values are as indicated unless **** p < 0.0001 by two-way ANOVA followed by Bonferroni’s multiple comparisons test ( d , e , g ). Source data are provided as a Source Data file.

Article Snippet: Twelve-week-old mice were acclimatized to single housing with HFD and intraperitoneal (i.p.) injection (PBS) for three days, followed by three daily leptin injections (2 mg/kg body weight, R&D systems; catalog 498-OB-05M) 1 h before the onset of dark cycle as described .

Techniques: Immunofluorescence, Staining, Injection, Quantitation Assay, Real-time Polymerase Chain Reaction, Western Blot