Drug tolerance requires m1G in mt-tRNA. (A-C) Illustration (A) of growth competition assays using GFP-labelled OCI-AML2 (B) and MOLM13 (C) control (sgCtr) and KO cells (sg2, sg4). (D) Frequency of apoptotic cells in populations of OCI-AML2 and MOLM13 control (sgCtr) and KO (sg2, sg4) cells using Annexin V labelling 3 to 5 days after cell sorting. (E-F) Quantification of mutated OCI-AML2 (E) and MOLM13 (F) cells (green line) overlaid with the frequency of viable TRMT5-KO (sg2) (black dots) and control (sgCtr) (white dots) cells over time. (G-H) Representative bright field image (G) and colony-forming efficiency assays (H) of OCI-AML2 control (sgCtr) and KO (sg2, sg4) cells in the presence of increasing concentrations of AraC. (I-L) Log2 FC of survival of drug-tolerant OCI-AML2 (I-K) and drug-sensitive MOLM13 (L) control (sgCtr) and TRMT5-KO (sg2, sg4) cells at the indicated time points. (M-N) Growth competition assay (M) and relative number of apoptotic cells after cytarabine (AraC) treatment (N) in Ctr and rescued TRMT5 KO cells. Cells were infected with a scramble (scr) or TRMT5 targeting gRNA (sg2). KO cells were rescued by reexpressing TRMT5 WT, catalytic dead (MUT) or lacking the MTS constructs. Control cells (sg2 and scr) were rescued with an EV control. Dunnett multiple comparisons test (panels D and N). Two-way analysis of variance multiple comparisons test (panels I-L).
