description: Ribosomal RNA depleted RNA was isolated, purified by using RiboMinus™ Eukaryote Kit (Invitrogen, #A10837-08) and chemically shredded into ~100-nt fragments by using RNA fragmentation reagents (Invitrogen, #AM8740). RNA fragments (2000 ng) were denatured at 95˚C for 3 min and incubated with 20 µl of Magna ChIP Protein A+G Magnetic Beads (Millipore, #2923270) conjugated to anti‑m6A antibody (2.5mg, Synaptic Systems, # 202003) or rabbit control IgG (Cell Signaling Technology) in 1X IPP buffer (15 mM NaCl, 10 mM Tris-HCl and 0.1% NP-40) with rotation at 4˚C for 4 h. The beads were washed twice with 1X IPP buffer, twice with low‑salt buffer (50 mM NaCl, 10 mM Tris-HCl and 0.1% NP-40), twice with high-salt buffer (500 mM NaCl, 10 mM Tris-HCl and 0.1% NP-40) and once with 1X IPP buffer. RNA was eluted from the beads with RLT buffer and purified through Qiagen RNeasy columns (Qiagen, #74104) according to the manufacturer's recommendation. RNA fragments were purified from the eluates with RNA Clean and Concentrator (Zymo) and used to construct libraries with TruSeq Stranded mRNA Library Prep Kit (Illumina, San Diego, CA). Sequencing was carried out on Illumina HiSeq 4000 according to the manufacturer's instructions with single-end 50-bp read length.
attributes.tag: strain
attributes.value: BALB/c
attributes.tag: isolate
attributes.value: MeRIP-Seq-Ctrl-1
attributes.tag: breed
attributes.value: not applicable
attributes.tag: cultivar
attributes.value: not applicable
attributes.tag: ecotype
attributes.value: not applicable
attributes.tag: age
attributes.value: not applicable
attributes.tag: dev_stage
attributes.value: not applicable
attributes.tag: sex
attributes.value: not applicable
attributes.tag: tissue
attributes.value: Intetinal epithelial cells
attributes.tag: cell_line
attributes.value: IEC4.1
attributes.tag: cell_type
attributes.value: Intestinal epithelial cells
attributes.tag: sample_type
attributes.value: Cell culture
attributes.tag: treatment
attributes.value: The IEC 4.1 murine intestinal epithelial cells were grown to 80% confluence for two groups: non-infected control (Ctrl) and Cryptosporidium parvum (CP)-infected group. Cells for CP infection were exposed to C. parvum infection for 24h. Cells were then collected and RNA isolated for m6A-MeRIP-Seq analysis.
attributes.tag: BioSampleModel
attributes.value: Model organism or animal