organism: Homo sapiens
description: Protocols: Human melanoma cell lines CHL-1 (WT), MEL224 (NRAS Q61R ) and A375 (BRAF V600E ) were transfected with expression vectors for GLI1, GLI2 or GLI3 proteins in two independent experiments. Non-transfected lines were used as controls. Briefly, 200 000 cells were seeded in a Ø10 cm cell culture dish and transfected the next day with 5 g of plasmid DNA, using the X-fect reagent (Clontech) and following the manufacturer's instructions. 48 hours post transfection, total RNA was isolated with Agilent Absolutely RNA miRNA Kit. The quality of the isolated RNA was checked on agarose gel, and concentrations of RNA were measured on NanoPhotometer N60 instrument (Implen, Munich, Germany). Sequencing libraries were generated and sequenced by DNA Link Company. The integrity of RNA samples was checked on Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). The instrument provided data on RNA concentration, electropherogram, rRNA subunit ratios and RNA integrity number (RIN). All samples had RIN ≥ 8.0 and a 28S:18S ratio ≥ 1.4. Total amount of 1 ug of RNA was used. cDNA libraries were prepared using TruSeq mRNA library Kit.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-01-01
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-01-01
attributes.tag: External Id
attributes.value: SAMEA110321872
attributes.tag: INSDC center alias
attributes.value: Garvan Institute of Medical Research
attributes.tag: INSDC center name
attributes.value: Garvan Institute of Medical Research
attributes.tag: INSDC first public
attributes.value: 2023-01-01T00:28:16Z
attributes.tag: INSDC last update
attributes.value: 2023-01-01T00:28:16Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-11936:Sample 7
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_line
attributes.value: A375
attributes.tag: cell_type
attributes.value: epithelial cell
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: melanoma
attributes.tag: genotype
attributes.value: GLI3 overexpression
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: skin
attributes.tag: sample name
attributes.value: E-MTAB-11936:Sample 7