quantifications of western blots with imagej ncbi

Quantifications of western blots with imagej ncbi license#
Quantifications of western blots with imagej ncbi plus#

d, A volcano plot representation of the microarray of miRNAs shown in Fig. Bar graphs represent a linear scale and error bars represent s.d. Data from biological triplicates are shown. b, c, qRT–PCR ( b) and western blot ( c) quantifications of METT元 upon transduction with two independent shRNAs targeting METT元 (METT元 KD1 and METT元 KD2) in MDA-MB-231 cells. A schematic representation of a pre-miRNA and a pri-miRNA is shown on the right. The magnitude of the over/underrepresentation is represented by the linear-scale heat map on the left. Yellow represents overrepresentation, and blue depicts underrepresentation of the motif.

Quantifications of western blots with imagej ncbi plus#

FIRE motif discovery analysis of pre-miRNAs and pri-miRNA sequences, as well as random sequences of the same length, reveals overrepresentation of the METT元 motif in pri-miRNAs sequences (containing pre-miRNAs plus adjacent 100 bp) but not in pre-miRNAs. Cell Stem Cell 15, 707–719 (2014)Ī, Unbiased search for cis-regulatory elements using the FIRE algorithm. m 6A RNA modification controls cell fate transition in mammalian embryonic stem cells. A sensitive non-radioactive northern blot method to detect small RNAs. Fiji: an open-source platform for biological-image analysis. Mapping in vivo protein–RNA interactions at single-nucleotide resolution from HITS-CLIP data. An integrated ChIP-seq analysis platform with customizable workflows. Fast gapped-read alignment with Bowtie 2. Endogenous human microRNAs that suppress breast cancer metastasis.

Quantifications of western blots with imagej ncbi license#

Beyond secondary structure: primary-sequence determinants license pri-miRNA hairpins for processing. In vitro and In vivo assays for the activity of Drosha complex. Detection of nonneutral substitution rates on mammalian phylogenies. Argonaute HITS-CLIP decodes microRNA–mRNA interaction maps. Structure prediction and phylogenetic analysis of a functionally diverse family of proteins homologous to the MT-A70 subunit of the human mRNA:m 6A methyltransferase. Integrative Genomics Viewer (IGV): high-performance genomics data visualization and exploration. N 6-methyladenosine residues in an intron-specific region of prolactin pre-mRNA. Effect of undermethylation on mRNA cytoplasmic appearance and half-life. Mapping and significance of the mRNA methylome. Purification and cDNA cloning of the AdoMet-binding subunit of the human mRNA (N 6-adenosine)-methyltransferase. Comprehensive analysis of mRNA methylation reveals enrichment in 3′ UTRs and near stop codons. Topology of the human and mouse m 6A RNA methylomes revealed by m 6A-seq. Nucleotide sequences at the N 6-methyladenosine sites of HeLa cell messenger ribonucleic acid. A universal framework for regulatory element discovery across all genomes and data types. Molecular basis for the recognition of primary microRNAs by the Drosha–DGCR8 complex. melanogaster homolog are required for miRNA biogenesis. The human DiGeorge syndrome critical region gene 8 and Its D. The Drosha–DGCR8 complex in primary microRNA processing. The Microprocessor complex mediates the genesis of microRNAs. Processing of primary microRNAs by the Microprocessor complex. Our findings reveal that the m 6A mark acts as a key post-transcriptional modification that promotes the initiation of miRNA biogenesis.ĭenli, A. Finally, gain-of-function experiments revealed that METT元 is sufficient to enhance miRNA maturation in a global and non-cell-type-specific manner. In vitro processing reactions confirmed the sufficiency of the N 6-methyladenosine (m 6A) mark in promoting pri-miRNA processing. Consistent with this, METT元 depletion reduced the binding of DGCR8 to pri-miRNAs and resulted in the global reduction of mature miRNAs and concomitant accumulation of unprocessed pri-miRNAs. Here we find in mammalian cells that methyltransferase-like 3 (METT元) methylates pri-miRNAs, marking them for recognition and processing by DGCR8. While the mechanisms underlying pri-miRNA processing have been determined, the mechanism by which DGCR8 recognizes and binds pri-miRNAs, as opposed to other secondary structures present in transcripts, is not understood. This initial event requires recognition of the junction between the stem and the flanking single-stranded RNA of the pri-miRNA hairpin by DGCR8 followed by recruitment of DROSHA, which cleaves the RNA duplex to yield the pre-miRNA product 5. The first step in the biogenesis of microRNAs is the processing of primary microRNAs (pri-miRNAs) by the microprocessor complex, composed of the RNA-binding protein DGCR8 and the type III RNase DROSHA 1, 2, 3, 4.