Fig 1: Genetic inhibition of mitochondrial translation suppresses GSC growth, recapitulating Q/D effects(A) Competition assay in VIPI cells transduced with viruses expressing sgRNAs for target genes. Data were normalized to day 1. n = 4 biological replicates, mean ± SD.(B) Percentage of insertions or deletions (indels) analyzed by decomposition (tracking of indels by decomposition [TIDE]) analysis in Cas9-expressing COMI and VIPI cells following lentiviral transduction of the sgRNAs selected in (A) (sgTUFM_1 and sgMRPS18A_1). n = 3–4 biological replicates, mean ± SD.(C) Effects of TUFM and MRPS18A knockout on TUFM, MRPS18A, and β-tubulin proteins in COMI and VIPI cells, as assayed by immunoblotting. One representative result is shown; n = 2 biological replicates.(D) Effects of TUFM and MRPS18A knockout on COX1, COX4, and β-tubulin proteins in COMI and VIPI cells, as assayed by immunoblotting. One representative result is shown; n = 2 biological replicates.(E) Effects of TUFM and MRPS18A knockout on COMI and VIPI cells grown in suspension. Shown are example images from days 0, 4, 10, and 15; scale bar, 200 μm.(F) Sphere area measured over the course of the 15-day experiment. The data were normalized to day 0. n = 15 technical replicates, mean ± SEM. One representative experiment is shown; n = 3 biological replicates.(G) Effects of TUFM and MRPS18A knockout on COMI and VIPI proliferation when grown as adherent cultures. The data were normalized to day 0. n = 3 biological replicates; mean ± SD.(H) Quantification of the number of spheres greater than 100 μm for COMI and VIPI cells; n = 20 technical replicates; mean ± SEM. ∗∗∗∗p < 0.0001, unpaired two-tailed t test. One representative result is shown; n = 3 biological replicates.