Metrics for growth responses of MDA-MB-231 and HME-1 cell lines to LINCS Pilot Phase TransCenter Project kinase inhibitors - Dataset (ID:20216)
|HMS Dataset ID:||20216|
|Dataset Title:||Metrics for growth responses of MDA-MB-231 and HME-1 cell lines to LINCS Pilot Phase TransCenter Project kinase inhibitors|
|Screening Lab Investigator:||Mario Niepel|
|Screening Principal Investigator:||Peter Sorger|
|Assay Description:||This dataset presents calculated metrics (IC50, GI50, EC50, Hill coefficient and Einf) for the data in HMS-LINCS dataset #20215. The data in 20215 are measures of the sensitivities of two cell lines to the LINCS Pilot Phase TransCenter kinase inhibitors -- cells were treated with single drugs and cell number was measured after three days of drug exposure.|
1. Cells were plated at 2000 cells per well in 4x 384 well plates per cell line and grown for 24 hours.|
2. Three plates per cell line were treated with single drugs using the HP D300 drug dispenser.
3. Immediately after treatment the untreated fourth plate for each cell line was fixed and stained by adding 20 μl of fixative containing formaldehyde solution and Hoechst 33342 for a final concentration of 3% formaldehyde and 250 ng/ml Hoechst (no wash step). This plate was sealed and kept at 4C until day 3 of the assay.
4. At day 3 the remaining plates were fixed, stained, and sealed as described in step 3.
5. After 1 hour all plates were scanned with a PE Operetta high throughput plate scanner (10x WD lens with excitation peak at 380nm and emission peak at 445nm).
6. Nuclei were counted using the Columbus software (module: 'Find Nuclei'; Method B; default parameters).
7. The results were normalized by plate and the three replicates were averaged to yield the average relative cell count and the relative growth for each cell line, drug and concentration.
8. Results at different concentration for a given drug and cell line are fitted with a sigmoidal curve to extract the IC50, GI50, EC50, Hill coefficient and Einf (see Fallahi-Sichani et al. Nat Chem Biol (2013) 9, p.708–714). A null value is set for cases where IC50, GI50 cannot be interpolated.
|Assay Protocol Reference:||Fallahi-Sichani M, Honarnejad, S Heiser, LM, Gray JW and Sorger PK. Metrics other than potency reveal systematic variation in responses to cancer drugs. Nature Chemical Biology, 2013. doi:10.1038/nchembio.1337. PMID: 24013279.|
|HMS Dataset Type:||Analysis|
|Date Publicly Available:||2014-12-10|
|Most Recent Update:||2016-09-09|