8 protocols
AccessionNameType
 
P-MTAB-33585
array scanning protocol
mRNA profiling was carried out on Illumina HumanHT-12 Expression BeadChips
P-MTAB-33584
nucleic acid hybridization to array protocol
mRNA profiling was carried out on Illumina HumanHT-12 Expression BeadChips
P-MTAB-33583
nucleic acid labeling protocol
mRNA profiling was carried out on Illumina HumanHT-12 Expression BeadChips
P-MTAB-33581
treatment protocol
NHEK Keratinocytes were transfectred in 24-well tissue culture plates using 0.8 ul of siPORT NeoFX Transfection agent (Life technologies) and 50000 cells in 0.6 ml medium (Keratinocyte-SFM medium kit, Invitrogen). Keratinocytes were transfected for 24h and stimulated either with 10ng/ml IFN-gamma (R&D)for 48h. Transfections were performed at concentrations of 30 nM of pre-miRNAs (Pre-miR miRNA Precursors Molecules hsa-miR-146a and Negative Control #1, Life technologies)
P-MTAB-33582
nucleic acid extraction protocol
miRNAeasy Mini Kit (Qiagen) was used to purify RNA from primary keratinocytes. The concentration and quality of RNA was assessed with NanoDrop ND-1000 spectrophotometer and Agilent RNA 6000 Nano Kit on Agilent 2100 Bioanalyzer.
P-MTAB-33579
growth protocol
Pooled Normal Human Epidermal Keratinocytes (NHEK) from adult healthy donors (Promocell) were cultured in serum free KC medium (Keratinocyte-SFM; Invitrogen) supplemented with 5 ng/mL human recombinant epidermal growth factor, 50 mg/mL bovine pituitary extract in a humidified atmosphere containing 5% CO2 at 37 deg C. Medium was changed every second day, and cells between passages 2 and 5 were used in all experiments.
P-MTAB-33580
treatment protocol
NHEK Keratinocytes were transfectred in 24-well tissue culture plates using 0.8 ul of siPORT NeoFX Transfection agent (Life technologies) and 50000 cells in 0.6 ml medium (Keratinocyte-SFM medium kit, Invitrogen). Keratinocytes were transfected for 24h and stimulated with 25 ng/ml TNF-alpha (both from R&D)for 48h. Transfections were performed at concentrations of 30 nM of pre-miRNAs (Pre-miR miRNA Precursors Molecules hsa-miR-146a and Negative Control #1, Life technologies)