2025-06-20
ENA first public
ERC000011
ENA-CHECKLIST
Department of Ophthalmology, Zibo Center Hospital
INSDC center name
public
INSDC status
ERS24832636
SRA accession
E-MTAB-15209:hRMECs with M1_2
Submitter Id
ArrayExpress
broker name
hRMEC
cell line
blood vessel endothelial cell
cell type
not collected
collection date
human
common name
Protocols: Human microglial clone 3 (HMC3) cells (Cat. No. CRL-3304) and primary renal mixed epithelial cells (hRMECs) (Cat. No. PCS-400-012) were purchased from ATCC. HMC3 cells were treated with 1 μg/mL LPS (Cat. No. HY-D1056, MCE, China) or 20 ng/mL IL-4 (Cat. No. HY-P70445, MCE) for 24 hs. HMC3 and hRMECs were co-cultured using Corning 12 mm Transwell with 0.4 μm pore polycarbonate membrane insert (Cat. No. 3401, Corning, USA) for 48 hs after induction.  Isolated RNA was confirmed RNA purity (A260/A280 ratio should be ~2.0) and integrity (RIN > 7) to ensure high-quality starting material. Use VAHTS mRNA Capture Beads (Vazyme) to capture polyadenylated mRNA, leaving out other RNA types. Convert the enriched or depleted RNA to complementary DNA (cDNA) through reverse transcription. Perform first-strand synthesis using reverse transcriptase and random primers. Follow with second-strand synthesis to create double-stranded cDNA. Perform first-strand synthesis using reverse transcriptase and random primers. Follow with second-strand synthesis to create double-stranded cDNA. Repair the ends of cDNA fragments and add a single \"A\" nucleotide at the 3' end to prevent self-ligation.
description
adult
developmental stage
Null syndrome
disease
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
retinal ganglion
organism part
Homo sapiens
scientific_name
hRMECs with M1_2
title