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Image Search Results
Journal: bioRxiv
Article Title: Whole-genome analysis of noncoding genetic variations identifies multigranular regulatory element perturbations associated with Hirschsprung disease
doi: 10.1101/2020.04.08.032045
Figure Lengend Snippet: Functional impacts of a HSCR-associated SNP (rs2435357) and the deletion of a novel S-HSCR enhancer on RET expression. ( a ) ATAC-seq and ChIP-seq profiles of hPSC and hNC in the intron 1 of RET show that rs2435357 is residing in a hNC-specific ATAC-seq peak. ( b ) Location of rs2435357 in the RET gene locus and in the sgRNA used for CRISPR/Cas9-mediated HDR for editing the C allele to the HSCR-associated risk allele T . The electrographs of Sanger sequencing show the successful introduction of the risk allele at rs2435357 in the UE-rs2435357 hPSC line. ( c ) Differentiation strategy to generate human neural crest (hNC) and neuronal progenitor (hNP), and immunostaining of SOX10 and TUJ1 in hNC and hNP of the control and the mutant (UE-rs2435357) lines. Scale bars: (hNC): 100 μ m; (hNP): 200 μ m. RT-qPCR analysis showing the comparable ELAVL4 expression level in hNP in the control ( n =5) and the mutant (UE-rs2435357) ( n =3) lines. t- test, ns : not significant. ( d ) RT-qPCR analysis showing RET expression in the hPSC and hNP stages of the control ( n =5) and the mutant (UE-rs2435357) ( n =3). t- test, ns : not significant. ( e ) Publicly available Hi-C data from GM12878 cells are shown using the RET locus as anchor. The putative enhancer in intron 1 of RASGEF1A is marked in yellow. ( f ) ATAC-seq and ChIP-seq data from hPSC and hNC at the RASGEF1A intron 1 locus. ( g ) The design of sgRNAs used for the CRISPR/Cas9 system for deleting the DNA fragment in RASGEF1A intron 1. Genotyping reveals the specific deletion of RASGEF1A intron 1 in UE-RASGEF1A-int1-KO hPSC line. WT: wildtype; KO: knockout. ( h ) Immunostaining of SOX10, TUJ1 and HU in hNC and hNP of the control and the mutant (UE-rs2435357) lines, respectively. Scale bars: (hNC): 100 μ m; (hNP): 200 μ m. ( i ) RT-qPCR reveals the expression level of RET in the hPSC and hNP stages of the control ( n =4-5) and the mutant (RASGEF1A-int1-KO) ( n =6-7). t- test, ns : not significant.
Article Snippet: Fixed cells were blocked with blocking solution (1% BSA, 0.1% Triton X-100 in PBS) at room temperature for 1 h. The blocked cells were then incubated with primary antibodies (
Techniques: Functional Assay, Expressing, ChIP-sequencing, CRISPR, Sequencing, Immunostaining, Mutagenesis, Quantitative RT-PCR, Hi-C, Knock-Out
Journal: Hearing research
Article Title: Culture media-based selection of endothelial cells, pericytes, and perivascular-resident macrophage-like melanocytes from the young mouse vestibular system
doi: 10.1016/j.heares.2016.12.012
Figure Lengend Snippet: Antibodies applied
Article Snippet: SOX10 ,
Techniques: