Journal: Nature Communications
Article Title: Acute depletion of BRG1 reveals its primary function as an activator of transcription
doi: 10.1038/s41467-024-48911-z
Figure Lengend Snippet: a Western blotting shows efficient depletion of BRG1 by Auxin treatment for 6, 10, and 24 h. Primary CD4 + T cells were isolated from BRG1-AID mice or wild-type mice. Following transduction of the cells with osTIR1 expression viral particles for 24 h, the cells were treated with Auxin for the times indicated on the top (6, 10, and 24 h). WT indicates the cells isolated from wildtype B6 mice; AID indicates the cells isolated from Brg1-AID knock-in B6 mice. Beta-actin and histone H3 were used as protein loading controls (repeated ≥ 2 times independently with similar results; uncropped scans of Fig. 1a are supplied as Source Data files). b A genome browser snapshot showing the BRG1 ChIC-seq signals in wild-type cells and BRG1-depleted cells. The cells were treated with Auxin for 10 h as described in Panel ( a ). Blue tracks: two replicates for wild-type cells; Red tracks: two panels for BRG1-depleted cells. c MA plot illustrating the genome-wide reduction in BRG1 binding (data from Panel ( b ) following 10 h of BRG1 depletion. Significantly altered BRG1 peaks are indicated, with decreases in red and increases in blue, from a total of 29,986 peaks. Significance was determined using a two-sided ROTS statistical test, with a cutoff of P < 0.05 and fold-change > 1.3. The P-values were calculated using the two-sided ROTS method, optimizing the reproducibility of feature ranking. d Bar plot showing the number of the BRG1 peaks at TSS, enhancers, and other regions in the wild-type CD4 + T cells. Note that enhancers are defined by the H3K27ac ChIC-seq data. e Bar plot showing the fractions of decreased BRG1 peaks at TSS, enhancers, and other regions in CD4 + T cells after BRG1 depletion for 10 h.
Article Snippet: The osTIR1 expression vector was obtained from Addgene (Plasmid #86233) .
Techniques: Western Blot, Isolation, Transduction, Expressing, Knock-In, Genome Wide, Binding Assay
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