Icons of wild variety (R722), heterozygous Epha2-Q722 (R722Q) and homozygous Epha2-Q722 (Q722) alleles utilizing three

Icons of wild variety (R722), heterozygous Epha2-Q722 (R722Q) and homozygous Epha2-Q722 (Q722) alleles utilizing three exon-13 primers (Table S1) AB928 Technical Information indicated by arrows within the schematic below. (B) PCR amplicons of wild sort (+/+), heterozygous Epha2-indel722 (+/indel722), and homozygous Epha2-indel722 (indel722) alleles using exon-13 flanking primers. Figure S2. RNA-seq information differential expression evaluation. Triplicate samples from wild-type (WT), Epha2-mutant (Q722, indel722), and Epha2-null lenses (P7) mostly cluster independently for all dysregulated genes (A). Complete heat-map of Figure 8A displays FC of every single gene relative to WT in every single Epha2 genotype tested (B). Figure S3. Gene ontogeny (GO) analysis of the combined upregulated genes from Epha2-mutant (Q722, indel722) and Epha2-null lenses (P7). Table S1. Primer sequences employed for PCR-amplification and Sanger sequencing of Epha2. Table S2. Primary antibodies employed for confocal microscopy, immunoprecipitation, and immunoblotting. Table S3. Differentially regulated genes (fold-change FC two, false discovery rate FDR 0.05) within the Epha2-Q722 lens (P7). Table S4. Differentially regulated genes (fold-change FC two, false discovery rate FDR 0.05) within the Epha2-indel722 lens (P7). Table S5. Differentially regulated genes (fold-change FC 2, false discovery price FDR 0.05) in the Epha2-null lens (P7). Author Contributions: All authors created substantial contributions to the perform and each has read and authorized the submitted version of your manuscript. Conceptualization Y.Z. plus a.S.; Methodology, Y.Z., P.A.R. along with a.S.; Validation, Y.Z. and P.A.R.; Formal Analysis, Y.Z. and P.A.R.; Investigation, Y.Z. and T.M.B.; Resources, P.A.R.; Data Curation, Y.Z. and P.A.R.; Writing–Original Draft Preparation, A.S.; Writing–Review and Editing, Y.Z., T.M.B., P.A.R. along with a.S.; Visualization, Y.Z., T.M.B. and P.A.R.; Supervision, A.S.; Project Administration, A.S.; Funding Acquisition, A.S. All authors have study and agreed for the published version of your manuscript. Funding: This perform was supported by NIH/NEI grants EY023549, EY028899 (to A.S.) and EY02687 (Core Grant for Vision Research) and an unrestricted grant for the Department of Ophthalmology and Visual Sciences from Research to prevent Blindness (RPB). GTAC is supported in part by NIH Grants P30 CA91842 and UL1TR002345. Institutional Assessment Board Statement: The Institutional Animal Care and Use Committee (IACUC) at Washington University approved all mouse procedures (Protocol No. 20190175) in compliance with all the Institute for Laboratory Animal Study (ILAR) guidelines. Data Availability Statement: RNA-seq data files have been deposited in the Gene Expression Omnibus (GEO) database below accession no. GSE181358. Acknowledgments: We thank B. McMahan and G. Ling for eye histology assistance, M. Casey for assist with gene-targeting design, and employees in the Genome Technology Access Center (GTAC), Genetic Engineering and iPSC Center (GEiC), and Mouse ES Cell Core facility at Washington University in St. Louis for help with RNA-sequencing and generation of gene-targeted mice. Conflicts of Interest: The authors declare no conflict of interest.
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