Journal: Redox Biology
Article Title: IDO1 improves postischemic neovascularization in aged mice by boosting endothelial NAD + de novo synthesis and curbing endothelial senescence
doi: 10.1016/j.redox.2025.103695
Figure Lengend Snippet: IL-17A/F represses ECs Ido1 transactivation and NAD + biosynthesis, and facilitates ECs senescence in the ischemic hindlimb muscle of young mice. A) Schematic diagram for testing methods was indicated. (B) Bar plot of the most prominent categories by KEGG pathway (left) and GO analysis (right). (C) Western blot analysis and quantification of IDO1 in hindlimb muscle ECs 7 days after HLI. PBS group was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗∗ P < 0.01, ∗∗∗ P < 0.001. (D) RT-qPCR quantification of Ido1 in hindlimb muscle ECs 7 days after HLI. PBS group was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗ P < 0.05. (E) Western blot analysis and quantification of IDO1 and P16 in hindlimb muscle ECs 7 days after HLI. Con group was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗∗ P < 0.01, ∗∗∗∗ P < 0.0001, ns: not significant. (F–I) RT-qPCR quantification of Ido1 (F), p16 (G) , p21 (H) and Lmnb1 (I) in hindlimb muscle ECs 7 days after HLI. Con group was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001, ns: not significant. (J–L) ROS (J), NAD + (K) and NO (L) levels quantification in in hindlimb muscle ECs 7 days after HLI. Data of Con group in J was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001, ns: not significant. (M) Venn diagram of IDO1 transcriptional factors between mouse and human. (N) Western blot analysis and quantification of P-CREB and total-CREB in hindlimb muscle ECs 7 days after HLI. Young group was set as 1. Unpaired two-tail t -test, n = 3 per group, data are mean ± SEM, ∗∗∗ P < 0.001. (O, P) Western blot analysis and quantification of P-CREB, total-CREB (T-CREB) (O), IDO1(P) and P16 (P) in hindlimb muscle ECs 7 days after HLI. Con group was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001, ns: not significant. (Q–T) RT-qPCR quantification of Ido1 (Q), p16 (R) , p21 (S) and Lmnb1 (T) in hindlimb muscle ECs 7 days after HLI. Con group was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ns: not significant. (U–W) ROS (U), NAD + (V) and NO (W) levels quantification in in hindlimb muscle ECs 7 days after HLI. Data of Con group in U was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001, ns: not significant.
Article Snippet: Mouse IL-17A protein (Catalog #HY- P70753 ), mouse IL-17F protein (Catalog #HY- P72584 ), mouse IL-17A/F protein (Catalog #HY- P77707 ), mouse IDO1 protein (Catalog #HY- P72616 ), IgG (Catalog #HY-P99002), brodalumab (Catalog #HY-P9925, an anti-interleukin-17-receptor IgG2 monoclonal antibody), and 666–15 (Catalog #HY-101120, CREB phosphorylation inhibitor) were purchased from Med-ChemExpress.
Techniques: Western Blot, Comparison, Quantitative RT-PCR