Extended Data Fig. 8. Single-cell drug response upon long-term exposure to EGFR inhibitors.

a, Single-cell analyses from data set presented in Fig 7b. After 72 hrs afatinib (1μM), EKAREN5 showed constant, non-oscillatory basal ERK signal. Rare activity spikes in few cells were observed in PDO-NRAS^Q61H and BRAF^V600E(#3).

b, Single-cell drug response analysis after 8 days of afatinib treatment (1μM). ERK dynamics in EGFR-inhibited PDO-KRAS^G12C (63 cells in 6 organoids) and PDO-NRAS^Q61H (70 cells in 5 organoids) are constant, non-oscillatory in nature. Depletion of the continuous oscillatory dynamics unmasked a hidden pattern of (EGFR-independent) rare activity spikes in few cells (one per ~30 hr (19 pulses observed in 571 hours of single-cell signaling evaluation). Never were two pulses observed in one single-cell derivation. Plots show all single cell analyses from multiple organoids or from a single representative organoid. Dotted line: to indicate that the two pulses are from separate cells.