For siRNA-mediated downregulation of Lamp3, Hela cells were transfected with LAMP3-specific siRNA (L-004716-01; ON-TARGETplus SMARTpool) or control siRNA (D-001810-01-05; ON-TARGETplus SMARTpool) using Dharmafect 1 transfection reagent (T-2004-01, Dharmacon [Thermo Scientific]; Lafayette, CO) according to the manufacture instructions. Following the initial 24 h transfection, cells were stimulated with IFN-alpha (Roferon-A, Hoffman-LaRoche) at the concentration of 1ng/ml and grown for 24 hr, then split, re-transfected with siRNA, and fresh medium with IFN was added. Cells were incubated for additional 24 or 48 h and collected at 48 hr and 72 hr of initial stimulation with IFN.
Extracted molecule
total RNA
Extraction protocol
RNA was isolated using RNAeasy kit (Qiagen).
Label
Cy3
Label protocol
amplification and labeling was performed using Ambion Illumina Total Prep RNA amplification kit
Hybridization protocol
Illumina BeadChip
Scan protocol
Illumina BeadChip
Description
HeLa24 5937290010_L
Data processing
AVG_Signal values generated by Bead studio software were imported into BRB Array Tools and median normalized before statistical comparisons
