Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)
Article Title: Blautia Coccoides is a Newly Identified Bacterium Increased by Leucine Deprivation and has a Novel Function in Improving Metabolic Disorders.
doi: 10.1002/advs.202309255
Figure Lengend Snippet: Figure 5. I3AA has beneficial effects on metabolic improvements in vitro and in vivo. A) Primary hepatocytes were incubated with indicate dose of I3AA for 48 h and then stimulated with 100 nM insulin for 20 min (n = 6 replicates per group). Western blot analysis of p-IR, p-AKT, and p-GSK3𝛽levels. The right panel is the densitometry analysis of the relative abundance of phosphorylated proteins normalized to their total protein levels. A.U.: arbitrary units. B–J) HFD mice were orally gavage with PBS or 10 mg kg−1 I3AA for 4 weeks (n = 6–7 biological replicates per group). B) Fed and fasting blood glucose levels. C) Fed and fasting serum insulin levels assayed by ELISA. D) HOMA-IR index. E) Glucose tolerance tests. The right panel is the AUC. F) Insulin tolerance tests (0.5 U kg−1). The right panel is the AUC. G) Total fat mass. H) The H&E staining of sWAT. Scale bars, 50 μm. The bottom panel is the frequency distribution of adipocyte cell size in sWAT and the box plot is average adipocyte diameter. I) Real-time PCR analysis of browning related genes (Ucp1, Pgc1𝛼, Prdm16, and Cidea) in sWAT. J) Western blot analysis of UCP1 protein levels. The bottom panel is the densitometry analysis of UCP1 protein levels. All values are expressed as the mean ± SEM. Statistical comparisons were carried out by unpaired two-tailed Student’s t-test; *p < 0.05 and **p < 0.01.
Article Snippet: Protein samples were subjected to concentration determination and immunoblot assay with the following antibodies: phosphorylated (p)-IR (Tyr 1150/1151; 3024S), total IR (3025), p-AKT (Ser 473; 9271S), total AKT (9272S), p-GSK3β (Ser 9; 9336S), and total GSK3β(9315S) from Cell Signaling Technology; AhR (17840-1-AP) and β–Actin (81115-1-RR) from Proteintech; FGF21 (ab171941) and UCP1 (ab209483) from Abcam.
Techniques: In Vitro, In Vivo, Incubation, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Real-time Polymerase Chain Reaction, Two Tailed Test