description: Protocols: DLD1 cells were cultured in Dulbecco's Modified Eagle Medium with GlutaMAX (Thermo Fisher) with 10% fetal bovine serum (Thermo Fisher). For mitotic synchronisation, cells were treated with 2 mM thymidine for 16 hrs, washed and released for 4 hrs, then treated with 9uM RO3306 for a further 16 hrs. Cells were washed and released for 45 minutes and mitotic cells were harvested by mitotic shakeoff. For PKCe inhibition, 500nM Blu577 or equivalent DMSO was added to the cells 2 hours before collection. In the case of mitotic cells, Blu577 was added to the cells 1.25 hours before mitotic release, and the release medium was also supplemented with Blu577. Cells were crosslinked with 140mJ/cm2 254nm UV irradiation, pelleted and snap frozen. The iCLIP protocol was followed to isolate RNA bound to the spliceosome and amplify a cDNA library as described in the manuscript. An anti-SERBP1 antibody (ab55993, Abcam) was used for immunoprecipitation. The iCLIP protocol was followed. Specific barcodes were used for each RT reaction, with random nucleotides also incorporated into cDNAs to distinguish between PCR duplication of cDNA products.
attributes.tag: ENA first public
attributes.value: 2021-10-05
attributes.tag: ENA last update
attributes.value: 2021-10-05
attributes.tag: External Id
attributes.value: SAMEA9637732
attributes.tag: INSDC center alias
attributes.value: The Francis Crick Institute
attributes.tag: INSDC center name
attributes.value: The Francis Crick Institute
attributes.tag: INSDC first public
attributes.value: 2021-10-05T08:19:09Z
attributes.tag: INSDC last update
attributes.value: 2021-10-05T08:19:09Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-10830:SERBP1_Asynchronous_DMSO_3
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: DLD1
attributes.tag: common name
attributes.value: human
attributes.tag: compound
attributes.value: DMSO
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: colorectal adenocarcinoma
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: immunoprecipitate
attributes.value: anti-SERB1
attributes.tag: irradiate
attributes.value: 140 millijoules per square centimetre of UV irradiation at 245 nm
attributes.tag: organism part
attributes.value: large intestine; colon
attributes.tag: sample name
attributes.value: E-MTAB-10830:SERBP1_Asynchronous_DMSO_3
attributes.tag: sex
attributes.value: male
attributes.tag: time
attributes.value: 2