Journal: Scientific Reports

Article Title: The 14th Ile residue is essential for Leptin function in regulating energy homeostasis in rat

doi: 10.1038/srep28508

Figure Lengend Snippet: ( A ) Daily food intake (Means ± SD) for WT (n = 5) and Lep ∆ I14/ ∆ I14 (n = 6) measured over 7–8 weeks. *P < 0.05 vs. controls. **P < 0.01 vs. controls. ***P < 0.001 vs. controls. ( B ) Body weight was measured over 9 weeks for WT (n = 5) and Lep ∆ I14/ ∆ I14 males (n = 6) and WT (n = 6) and Lep ∆ I14/ ∆ I14 females (n = 6). *P < 0.01 vs. controls. **P < 0.01 vs. controls. ***P < 0.001 vs. controls. ( C ) Serum INSULIN level in 8-wk-old male WT (n = 4) and Lep ∆ I14/ ∆ I14 rats (n = 5), and female WT (n = 4) and Lep ∆ I14/ ∆ I14 rats (n = 6). ***P < 0.001 vs. controls. ( D ) Male WT (n = 5) and Lep ∆ I14/ ∆ I14 rats (n = 6) at age of wk-16 were ip injected with D-glucose, and serum glucose levels were determined at 0, 30, 60, 90, and 120 min after administration. *P < 0.05 vs. controls. ( E ) Hyperphagia and obesity were rescued by subcutaneous infusion of recombinant LEP. Two individual female Lep ∆ I14/ ∆ I14 rats were treated with rat recombinant Lep via subcutaneous implantation of an Alzet osmotic minipump (200 μg/kg/day) for 7 days before switching LEP to saline, while the WT female controls (n = 3) were implanted with a saline-filled minipump. For Lep ∆ I14/ ∆ I14 rats, solid arrow indicates the start of LEP treatment; solid arrowhead indicates the switch of LEP to saline. For WT controls, open arrow indicates the start of minipump implantation; open arrowhead indicates withdrawal of minipump. Numbers near each body-weight point are the daily food intake (g) measured on the same day.

Article Snippet: Each Lep ∆ I14/ ∆ I14 rat was subcutaneously implanted an Alzet osmotic minipump (model 2001, Durect) filled with rat recombinant LEP (Genscript) at a dosage of 200 μg/kg/day (in sterile saline), while each WT rat was subcutaneously implanted an Alzet osmotic minipump (model 2001, Durect) filled with sterile saline as control.

Techniques: Injection, Recombinant, Saline

Journal: Scientific Reports

Article Title: The 14th Ile residue is essential for Leptin function in regulating energy homeostasis in rat

doi: 10.1038/srep28508

Figure Lengend Snippet: ( A ) Sanger-sequencing of the RT-PCR products showed that the Lep mRNA of Lep ∆ I14/ ∆ I14 rat had a deletion of 3 nucleotides ATC encoding an Ile residue. ( B ) Western blot showed that the mature LEP ∆I14 protein is stably expressed in the WAT of Lep ∆ I14/ ∆ I14 rats. Shown is one of three independent experiments. ( C ) ELISA showed that serum LEP ∆I14 in male Lep ∆ I14/ ∆ I14 rats (n = 5) is significantly increased compared to that of serum LEP WT in the male WT controls (n = 5). ( D ) Computer assimilation of LEP-LEPR interaction using available information from their human homologs: LEP (PDB number: 1AX8) and LEPR (PDB number: 3V6O). ( E ) STAT3 reporter assay. 293FT cells were treated with WT and mutant recombinant rat LEP proteins at different concentrations after transient transfection of pcDNA-Lepr, pRL-TK and pGL6-Stat3. Relative luciferase activity was determined by firefly luciferase light units normalized by that of renilla luciferase.

Article Snippet: Each Lep ∆ I14/ ∆ I14 rat was subcutaneously implanted an Alzet osmotic minipump (model 2001, Durect) filled with rat recombinant LEP (Genscript) at a dosage of 200 μg/kg/day (in sterile saline), while each WT rat was subcutaneously implanted an Alzet osmotic minipump (model 2001, Durect) filled with sterile saline as control.

Techniques: Sequencing, Reverse Transcription Polymerase Chain Reaction, Residue, Western Blot, Stable Transfection, Enzyme-linked Immunosorbent Assay, Reporter Assay, Mutagenesis, Recombinant, Transfection, Luciferase, Activity Assay