sirt2 sirna am16708  (Thermo Fisher)

Journal: Advanced Science

Article Title: Adult Mouse Kidney Stem Cells Orchestrate the De Novo Assembly of a Nephron via Sirt2‐Modulated Canonical Wnt/ β ‐Catenin Signaling

doi: 10.1002/advs.202104034

Figure Lengend Snippet: Sirt2 is required for dynamic change of canonical Wnt/ β ‐catenin signaling during kidney organoid development. A) Model of self‐organized kidney organoid development from single kidney stem cells. Scale bar, 100 µm. B) Western blot analysis of expression of β ‐catenin, GSK3 β , Sirt2, and podocin1/2 during kidney organoid development. GAPDH was used as loading control for quantitation. C–F) Quantification of gene expression for β ‐catenin, GSK3 β , Sirt2, and podocin1/2, respectively. G) Western blot analysis of Sirt2, β ‐catenin, and podocin1/2 in KSC‐Sirt2 wt or KSC‐Sirt2 knockdown (KD) kidney organoids. * p < 0.05 versus day 0.

Article Snippet: To knockdown Sirt2, mouse Sirt2 siRNA (Thermo Fisher Scientific, #AM16708) and negative control (Sigma, #SIC001‐10NMOL) were transfected into kidney stem cells (passage 66) with Lipofectamine 3000 transfection reagent (Thermo Fisher Scientific, #L3000008) at the final concentration of 5nM following the manufacture's instruction for 18 h. Then, transfected cells were cultured in Medium C for 48 h and used for kidney organoids study.

Techniques: Western Blot, Expressing, Control, Quantitation Assay, Gene Expression, Knockdown

Journal: Advanced Science

Article Title: Adult Mouse Kidney Stem Cells Orchestrate the De Novo Assembly of a Nephron via Sirt2‐Modulated Canonical Wnt/ β ‐Catenin Signaling

doi: 10.1002/advs.202104034

Figure Lengend Snippet: Schematic model of Sirt2‐mediated development of adult kidney stem cells‐derived kidney organoids. Adult kidney stem cells (KSCs) reside in the mouse kidney. KSCs were isolated using Sca1/Oct4 selection markers and labeled with mCherry and CD63‐GFP. KSCs formed self‐organized kidney organoid in culture mediated by Sirt2 expression. Sirt2 regulates GSK3 β activity leading to a dynamic change of canonical Wnt/ β ‐catenin signaling and promotes the formation of KSCs‐derived kidney organoid and nephron development.

Article Snippet: To knockdown Sirt2, mouse Sirt2 siRNA (Thermo Fisher Scientific, #AM16708) and negative control (Sigma, #SIC001‐10NMOL) were transfected into kidney stem cells (passage 66) with Lipofectamine 3000 transfection reagent (Thermo Fisher Scientific, #L3000008) at the final concentration of 5nM following the manufacture's instruction for 18 h. Then, transfected cells were cultured in Medium C for 48 h and used for kidney organoids study.

Techniques: Derivative Assay, Isolation, Selection, Labeling, Expressing, Activity Assay