1. Fig. 3. TKI sensitivity assessed by the MANO method.
TKI sensitivity assessed by the MANO method. (A) Ba/F3 cells expressing each of the 16 genes shown on the right were mixed and cultured in the presence of different concentrations of gefitinib, erlotinib, afatinib, osimertinib, rociletinib, crizotinib, alectinib, or puromycin. Bar code read numbers of the compound-treated cells were normalized to those of the dimethyl sulfoxide (DMSO)–treated mixture, and the relative viability (%) of each cell clone on day 5 is color-coded according to the indicated scheme. (B) Comparison of cell viability measured with alamarBlue cell viability assay and the MANO method for Ba/F3 cells with 10 EGFR mutants in (A). Each data point was normalized to vehicle-treated cells. Pearson’s correlation coefficient (r) was calculated as 0.89 (P < ). The low ratio area is magnified in the right panel. (C) Changes in the relative cell populations in the tumors of mice treated with TKIs. The MANO method was used to quantify the bar code read numbers of tumors in 10 erlotinib-treated and 10 vehicle-treated mice. The bar code number for each cell line was normalized to the total bar code numbers of the tumor on day 18, and the calculated number was subsequently used to determine the percentage contribution of each cell line to the tumors. The mean relative cell population (%) within the tumors is shown for mice treated with either vehicle (green circle) or erlotinib (orange circle). The blue and pink arrows represent decrease and increase in the relative cell populations, respectively, in the tumors treated with erlotinib compared to those treated with vehicle. Error bars denote SD.
Shinji Kohsaka et al., Sci Transl Med 2017;9:eaan6566
Published by AAAS