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TranscriptomicseIF4A3 - Modified iCLIP
The modified iCLIP protocol is based on the previously described protocol (Huppertz et al., 2014) with modifications that enable the definition of readthrough cDNAs. HeLa cells were crosslinked with 0.15mJ/cm2 254nm UV light. Protein G Dynabeads were used for immunoprecipitations (IP). For each IP, ...
ORGANISM(S): Homo sapiens 
2016-11-24 | E-MTAB-4000 | biostudies-arrayexpress
TranscriptomicsPTBP1 - Modified iCLIP
This experiments was performed in HeLa cells according to the iCLIP protocol with the following modifications: no antiRNase was used and the concentration of RNase I was 0.5 U/ml. In iCLIP4, the dephosphorylation step was omitted from the standard protocol. The rest of the protocol was identical to...
ORGANISM(S): Homo sapiens 
2016-11-24 | E-MTAB-5026 | biostudies-arrayexpress
TranscriptomicsModified iCLIP sequencing of HEK293 cells with antibody PTBP1
For modified iCLIP experiment, 4SU was used for crosslinking as described in published protocol (citation is bellow) and the RNase conditions were optimised to ensure efficient RNase I-dependent fragmentation. In detail, HEK293T cells were grown on 10 cm 2 dishes, incubated for 8 h with 100 M 4SU an...
ORGANISM(S): Homo sapiens 
2016-11-24 | E-MTAB-5027 | biostudies-arrayexpress
Transcriptomicsmodified iCLIP experiment with the addition of a 5'marker to identify read-through reads
We added an oligonucleotide to the 5′ end of RNA fragments —5′ marker— to differentiate between truncated and read–through cDNAs. By applying the protocol to PTBP1 and eIF4A3 iCLIP, we determined that the start sites of read–through cDNAs are enriched in adenosines, while the truncated cDNAs prefere...
ORGANISM(S): Homo sapiens 
2018-06-15 | E-MTAB-6927 | biostudies-arrayexpress
TranscriptomicseIFA3 - iCLIP
The exon junction complex is deposited at 24nt upstream of exon-exon junctions, but not at every junction. The core complex is comprosed of 4 proteins, eIF4A3, Magoh, Y14 and MLN51. Here we performed immunoprecipitation of Y14 with subsequent iCLIP of eIF4A3 in HeLa cells to identify the crosslink s...
ORGANISM(S): Homo sapiens 
2016-11-24 | E-MTAB-3618 | biostudies-arrayexpress
ProteomicsYBX3 is a versatile RNA-binding protein that controls amino acid import by regulating SLC mRNA abundance
Summary: Amino acids (AA) support protein synthesis, and thus cell growth and proliferation. Specialized transporters mediate AA exchange across membranes and their regulation is critical for AA homeostasis. Here, we report that the DNA- and RNA-binding protein YBX3 regulates the expression of AA t...
ORGANISM(S): Homo Sapiens (ncbitaxon:9606) 
2018-11-06 | MSV000083108 | MassIVE
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