I previously did some recombined cell enrichment curves using flow cytometry of mixed cells populations (containing both recombined and unrecombined cells) upon treatment with various selections (Blast, Puro, Hygro, Zeo), but my recent experience testing out the various plate readers got me in the mode of doing serial dilutions with various cell viability / (relative) cell counting assays, so I did that for our mostly commonly used Single Landing Pad HEK cells (LLP-Int-BFP-IRES-iCasp9-Blast) as well as our still unpublished Double Landing Pad HEK cells. Here are what those curves generally look like.
Note: The Double Landing Pad cells encode hygromycin resistance within the second landing pad, so the difference in hygromycin IC90’s in the SLP and DLP cells is expected. Once we have the plate reader purchased and back in the lab, I’ll probably do some other conditions (eg. unmodified 293Ts, recombined SLP cells, recombined DLP cells) for some of those common cell culture antibiotics.
Note 2: Also, now that I know the general effective concentrations of some of these chemotherapeutics (eg. cyclophosphamide, gemcitibine, vincristine; although I did completely overshoot the concentrations for gemcitibine), I’m somewhat set up to see how the IC90 changes when I overexpress various transgenes that should increase resistance to those drugs.