Journal: Molecular Medicine

Article Title: Exenatide regulates Th17/Treg balance via PI3K/Akt/FoxO1 pathway in db/db mice

doi: 10.1186/s10020-022-00574-6

Figure Lengend Snippet: Effects of exenatide on Th17/Treg proliferation in vitro. A The expression of GLP-1R in PBMCs from C57BL/6 J mice by western blot. Representative image and gray value analysis of GLP-1R from Control and palmitate (PA) groups. Data are presented as mean ± SEM of three independent experiments. ACTIN was used as a loading control. Naïve CD4 + T cells collected from C57BL/6 J mice were cultured under Th17/Treg-inducing conditions with palmitate (PA), exenatide plus palmitate (Exe + PA) and exenatide, palmitate plus FxoO1 inhibitor AS1842856 (Exe + PA + AS). B Representative flow cytometric plots of Th17 cells from different groups. C Statistical analysis of the percentage of Th17 cells in different groups. D Representative flow cytometric plots of Treg cells with CD4-FITC, CD25-PE and Foxp3 staining. E Statistical analysis of the percentage of Treg cells. Data are presented as mean ± SEM of three independent experiments. ** P < 0.01, * P < 0.05 vs control group; # P < 0.05 vs PA group; & P < 0.05 vs Exe + PA group; ns P > 0.05

Article Snippet: After blocking with 5% non-fat milk in Tris-buffered saline Tween-20 (TBST), the membranes were incubated with primary antibodies against IL-17 (#AO688, Abclonal Technology, China), Foxp3 (#GB11093, Servicebio, China), phosphoinositide 3-kinase (PI3K) (#bsm-33219m, Bioss, China), phospho (p)-PI3K (Tyr317, #bs-5570R, Bioss, China), protein kinase B (Akt) (#GB11689, Servicebio, China), p-Akt (Ser473, #AF0908, Affinity, USA), FoxO1 (#GB11286, Servicebio, China), p-FoxO1 (Thr24, #9464T, Cell Signaling Technology, USA) or GLP-1R (#CSB-PA009514YA01HU, Cusabio, China) at 4°C overnight.

Techniques: In Vitro, Expressing, Western Blot, Control, Cell Culture, Staining