Extended Data Fig. 5. CYPA is required for cellular activity.
a, Cellular CYPA protein levels determined by parallel reaction monitoring (PRM) mass spectrometry in NCI-H358 cells harboring doxycycline-inducible expression of low or high CYPA in the absence or presence of doxycycline (0.1 µg/ml) for 120 h. Bars indicate the mean of 3 biological replicates per group from one experiment. b, pERK levels (MSD) of CYPA high and CYPA low cells treated with RMC-7977 in the absence or presence of doxycycline (0.1 µg/ml) for 120 h. Datapoints show biological duplicates normalized to vehicle control. Representative data shown from one of 3 independent experiments. c, Cellular CYPA protein concentration in a panel of cell lines determined by PRM. Data 2 stable isotope labeled peptide standards (SIS) was averaged for each replicate. Intracellular µM concentration estimated assuming cell volume of 2 pl. Bars indicate the mean of biological duplicates. d, CYPA protein expression in cells (green bars) and corresponding xenograft tumours (blue bars) determined by PRM. 5 SIS peptides were used and data averaged for each biological replicate. Bars indicate the mean of biological duplicates for HPAC cells and triplicates for all others. e, PRISM screen, RMC-7977 sensitivity (AUC) by PPIA gene expression (RPKM). Each dot represents a cell line (n = 606). Two-sided Pearson correlation = −0.10 (p = 0.011).