Screen Status: Primary miRNA
Screen Overview: Primary Screen
Screener: Elza De Bruin (Downward Lab).
Extension:3365, Rm 206
Library: Human miRNA Mimics 384
Library Description: Qiagen library of 10 plates miRNA mimics stamped into 384 well plates
Library Supplier: Qiagen
Library Format: 3 plate(s) in 384 well format
Replicate Info: 3 replicates
Publications: Reduced NF1 expression confers resistance to EGFR inhibition in lung cancer.. de Bruin et al., 2014
Keywords: Erlotinib, Lung Cancer, Viability
Cell Line: PC-9

Screen Method

Day 1: Transfection of PC9 cells

PC9 cells were transfected using Dharmafect2 transfection reagent and cultured for 2 days.

Day 3: Addition of Erlotinib

For the drug treated plates, the culture media was aspirated and fresh media containining Erlotinib was added (final concentration 30nM). For control plates, fresh media only was added. The plates were then cultured for a further 3 days.

Day 6: Fix the plates with 80% EthOH and stain the plates with DAPI

Plates were fixed with 80% Ethanol and stained with DAPI.

Readout

Acumen Explorer eX3 laser scanning cytometer was used to determine cell number per well.