Figure 2. MTOR is an essential kinase in Colo699 cells.

An essential kinase screen was performed with the kinome shRNA lentiviral library as described in the Materials and Methods. In A, the count frequencies following Illumina sequencing of six MTOR targeting shRNAs is presented for Colo699 and 584-A2 cells. The findings show loss of five of the six MTOR shRNAs by 7 days of culture in Colo699 cells, but not 584-A2 cells. B, Colo699 cells were transduced with either a negative control shRNA targeting GFP, or one of two independent MTOR targeting shRNAs (MTOR 22 or MTOR 88). Cells were selected for resistance to puromycin and resulting colonies following 2 weeks of culture were stained with crystal violet and total colony area was quantified. C, The indicated cell lines and others noted in Table 1 were submitted to clonogenic growth assays (see Materials and Methods) with 0 – 1 μM AZD8055. After ~ 2 weeks, the colonies were fixed and stained with crystal violet and total colony area was quantified. The IC₅₀ values were calculated with the Prism software program and presented in Table 1.