Extended Data Fig. 3: In vivo validation and characterization of primary hits.
From: Disruption of the MSL complex inhibits tumour maintenance by exacerbating chromosomal instability
a, Growth kinetics of the indicated conditions in tumor maintenance assays performed as indicated in Fig. 1e. N = 4 biologically independent tumors. P-value from one-tailed Student’s t-test at the last time point. b, Quantification of gene editing efficiency of TDF cells of the indicated samples as assessed by Sanger sequencing and subsequent TIDE analysis https://tide.deskgen.com/52. Cntr: control cells expressing GFP-targeting sgRNAs. c, Schematic representation of the four MSL subunits and the mutants generated by genome editing. Arrowheads indicate the approximate location of sgRNAs used to generate truncated proteins lacking functional domains used in d, The characterized function of each domain is described in Supplementary Table 3. CC: coiled-coil. CD: chromo. CB: chromobarrel. d, Quantification of MSL loss-of-function in the indicated MSL-mutant cell populations generated using the sgRNAs indicated in c, H4K16ac levels were measured by quantitative immunofluorescence and used as a readout of MSL function. The effect of KAT8-KO is underestimated as many cells H4K16aclow died and detached from the plate prior to staining. Note that slight differences in the fraction of H4K16aclow cells across mutants may be due to differences in sgRNA activity. All mutants resulted in reduction of the histone mark. The number of analyzed cells in each condition is shown. The statistical significance of the differences compared to a control sgRNA are indicated by three asterisks (p < 0.0001, two-tailed Fisher’s test) e-f, Representative images (e) and quantification (f) of H4K16ac levels in TDF cells at the indicated times after induction of MSL1 knock-out. Values are average ± SEM from n = 3 biologically independent samples. P-value from one-way ANOVA. Scale bar: 20 µm. g, Quantification of gene editing in the indicated tumors by Sanger sequencing and subsequent TIDE analysis. Tumors showing less than 50% edited sequence were considered unedited. Note the varying editing efficiency across all tumors, indicating the presence of wild-type cells also in tumors classified as “edited”. Wild-type cells sustain tumor growth, leading to an underestimation of the effect induced by MSL disruption. Source data are provided.
