Journal: Nature Communications
Article Title: NCoR1 restrains thymic negative selection by repressing Bim expression to spare thymocytes undergoing positive selection
Figure Lengend Snippet: NCoR1 binds to the Bim promoter and represses Bim expression. a RNA-Seq analysis of differentially expressed genes in NCoR1-deficient (cKO) and wild-type (WT) thymocytes with or without activation in vitro. b , c Quantitative RT-PCR analysis of Bim mRNA expression in freshly sorted cKO ( n = 6) and WT ( n = 6) CD4 + CD8 + double-positive (DP) thymocytes ( b ), and in total cKO ( n = 4) and WT ( n = 4) thymocytes ( c ) after in vitro TCR stimulation for the indicated amounts of time. d Immunoblot analysis of NCoR1 and Bim expression levels in extracts of cKO and WT thymocytes with or without stimulation for the indicated amounts of time. e Quantification of NCoR1 and Bim expression levels in ( d ). f ChIP analysis of the Bim promoter by anti-NCoR1 in cKO and WT thymocytes with or without activation in vitro. IgG served as a negative control. g ChIP analysis of the Bim promoter in cKO and WT thymocytes using the indicted antibodies. The data are representative of two independent experiments ( b , c , f , g ). Statistical significance was analyzed using the two-tailed Student’s t test (* P
Article Snippet: After crosslinking with goat antibody to hamster immunoglobulin G (20 μg ml−1 ; 55397; MP Biomedicals) at various time points, the cells were lysed and subjected to SDS-PAGE.
Techniques: Expressing, RNA Sequencing Assay, Activation Assay, In Vitro, Quantitative RT-PCR, Chromatin Immunoprecipitation, Negative Control, Two Tailed Test