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The scar-6 elncRNA gene epigentically regulates expression of prozb via prdm14-PRC2 complex. (A) UCSC genome browser screenshot representing JASPER 2022 track with 12 bp deletion of scar-6 locus. (B) Bar plot representing ChIP-qPCR quantification in percentage input using prdm14 antibody as a target and mock as a control for scar-6 locus in wild type and scar-6gib007Δ12/Δ12 mutant zebrafish. Data from 3 independent biological replicates plotted as mean fold change ± standard deviation; ns, not significant, *p < 0.05 (two-tailed unpaired t-test). (C) Box and whiskers plot representing percentage methylation profile of proximal CpG island of scar-6 using targeted bisulfite sequencing of wild type and scar-6gib007Δ12/Δ12 mutant zebrafish per cytosine. Data from 4 independent biological replicates plotted with line at mean. (D) Bar plot representing fold enrichment of scar-6 elncRNA upon RNA-immunoprecipitation of prdm14 and Suz12 followed by qPCR of scar-6 elncRNA. The scar-6 elncRNA shows enrichment in the binding with prdm14 and Suz12. Data from 3 independent biological replicates plotted as mean fold change ± standard deviation; ***p-value ≤ 0.001, ****p < 0.0001 (two-tailed unpaired t-test). (E) Bar plot representing ChIP-qPCR quantification in percentage input using prdm14 antibody as a target and mock as a control for scar-6 locus in zebrafish injected with scar-6 elncRNA morpholino (scar-6 MO) and scrambled morpholino (scr MO). Data from 3 independent biological replicates plotted as mean fold change ± standard deviation; **p < 0.01, ***p < 0.001, ****p < 0.0001 (two-tailed unpaired t-test). (F) Schematic representation of prdm14-PCR-2 complex binding at scar-6 locus and being stabalized by the scar-6 elncRNA produced from the scar-6 elncRNA gene. Source data are available online for this figure.
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