Journal: Communications Biology
Article Title: Sirt6 -mediated epigenetic modification of DNA accessibility is essential for Pou2f3 -induced thymic tuft cell development
Figure Lengend Snippet: a , b scATAC-seq data of WT and Sirt6 -deficient mTECs are coembedded into a single UMAP visualization ( a ) and are displayed separately ( b ). Cells are stained with the mTEC subset defined before. c Relative proportion of mTEC clusters between WT and Sirt6 cKO mice. d Simple Western analysis of POU2F3 expression in sorted mTECs (CD45 − EpCAM + UEA-1 + Ly51 − ) from 4-week-old WT and Sirt6 cKO mice. e UMAP visualization of the gene expression of Pou2f3 in WT (left) and Sirt6 cKO (right) scATAC-seq. The red scale in the UMAP plot indicates the level of Pou2f3 gene activity in scATAC-seq f UMAP visualization of the motif accessibility score of Pou2f3 in WT (left) and Sirt6 cKO (right) scATAC-seq. The blue scale in the UMAP plot indicates the level of Pou2f3 motif accessibility in scATAC-seq analyzed by chromVAR. g , h Motif accessibility scores of Klf3 ( g ) and Pou2f3 ( h ) in Late Aire 2 cells and thymic tuft cells of WT (Orange) and Sirt6 cKO mice (green) were compared in a violin plot. WT Late Aire 2 cells: n = 272, Sirt6 cKO Late Aire 2 cells: n = 500, WT thymic tuft cells: n = 501, Sirt6 cKO thymic tuft cells: n = 150. i , j Chromatin accessibility peaks of overlapping genes ( i ) and tuft-associated genes ( j ) mentioned in Fig. in WT and Sirt6 -deficient thymic tuft cells are shown in aggregated scATAC-seq tracks. The arrow indicates the length and direction of the genes detected by scATAC-seq. The statistics was determined using Wilcoxon signed-rank test. * p < 0.05, ** p < 0.01, *** p < 0.005 and **** p < 0.001.
Article Snippet: The primary antibody, anti-POU2F3 (Prosci, 7795), was diluted with Antibody Diluent II at 1:40, while the anti-rabbit secondary HRP antibody (042-206; ProteinSimple) was used directly.
Techniques: Staining, Western Blot, Expressing, Activity Assay