Journal: eLife
Article Title: KRAB-zinc finger protein gene expansion in response to active retrotransposons in the murine lineage
doi: 10.7554/eLife.56337
Figure Lengend Snippet: ( A ) Differential gene expression in Chr2-cl and Chr4-cl KO ES cells. Significantly up- and downregulated genes ( adjusted p-value <0.05) are highlighted in red and green, respectively, KRAB-ZFP genes within the deleted clusters are shown in blue. ( B ) Correlation of TEs and gene deregulation. Plots show enrichment of TE groups within 100 kb of up- and downregulated genes relative to all genes. Significantly overrepresented LTR and LINE groups ( adjusted p-value <0.1) are highlighted in blue and red, respectively. ( C ) Schematic view of the downstream region of Chst1 where a 5’ truncated ETn insertion is located. ChIP-seq (Input subtracted from ChIP) data for overexpressed epitope-tagged Gm13051 (a Chr4-cl KRAB-ZFP) in F9 EC cells, and re-mapped KAP1 (GEO accession: GSM1406445) and H3K9me3 (GEO accession: GSM1327148) in WT ES cells are shown together with RNA-seq data from Chr4-cl WT and KO ES cells (mapped using Bowtie (-a -m 1 -- strata -v 2) to exclude reads that cannot be uniquely mapped). ( D ) RT-qPCR analysis of Chst1 mRNA expression in Chr4-cl WT and KO ES cells with or without the CRISPR/Cas9 deleted ETn insertion near Chst1 . Values represent mean expression (normalized to Gapdh) from three biological replicates per sample (each performed in three technical replicates) in arbitrary units. Error bars represent standard deviation and asterisks indicate significance (p<0.01, Student’s t-test). n.s.: not significant. ( E ) Mean coverage of ChIP-seq data (Input subtracted from ChIP) in Chr4-cl WT and KO ES cells over 127 full-length ETn insertions. The binding sites of the Chr4-cl KRAB-ZFPs Rex2 and Gm13051 are indicated by dashed lines.
Article Snippet: Cell line ( Mus musculus ) , F9 Embryonic carcinoma cells , ATCC , ATCC CRL-1720 , .
Techniques: Gene Expression, ChIP-sequencing, RNA Sequencing, Quantitative RT-PCR, Expressing, CRISPR, Standard Deviation, Binding Assay