Extended Data Fig. 4: Disruption of the MSL complex impairs the long-term proliferative capacity of cancer cells.
From: Disruption of the MSL complex inhibits tumour maintenance by exacerbating chromosomal instability
a, Quantification of the number of adherent, viable cells 8 d after induction of KAT8 knock-out with doxycycline (KAT8-KO) and in the corresponding uninduced cells. Values are average ± SEM from n = 3 biologically independent samples. P-value from one-tailed Student’s t-test. b, Images of TDF cells 4 d after transfection of synthetic crRNAs-trRNAs targeting KAT8 (KAT8-KO) or the GFP reporter (cntr-KO). Scale bar: 1 mm. c, Quantification of H4K16ac levels in adherent, alive cells and detached cells recovered from the medium 4 d after induction of KAT8 knock-out by doxycycline treatment. For boxplots, top, middle and bottom delimiters: 75th, 50th, 25th percentiles; top and bottom whiskers: 90th and 10th percentiles. N = 471 and 488 biologically independent adherent and detached cells, respectively. P-value from one-tailed Mann Whitney’s U-test. AU: arbitrary units. d, Limiting dilution clonogenic assays of the indicated TDF cells 11 d after plating. Representative images of 96-well plates in which 2-fold serial dilutions of cells were plated. External wells are not shown (see Methods). Yellow shows the IncuCyte cell mask detecting growing clones. Scale bar: 6 mm. e, Limiting dilution clonogenic assays using TDF cells. The percentage of wells containing at least one clone larger than 20 cells is plotted. Values are average ± SEM from n = 3 biologically independent samples. The knock-out efficiency, as assessed by quantification of H4K16aclow cells in each population is indicated. P-value from one-tailed Student’s t-test. f, Limiting dilution transplantation assay into NSG mice using cells from unedited or MSLLOF (MSL3-KO) tumors. The frequency of tumor-propagating cells (estimate with upper and lower limits) is indicated. P-value from χ2 test. g-h, Quantification (g) and representative images (h) of H4K16ac levels in HT-1080 cells at the indicated times after induction of MSL1 knock-out. Values are average ± SEM from n = 3 areas in the well for a total of over 800 cells per condition. P-value from two-tailed Fisher’s exact test. Scale bar: 100 µm. i, Viability phenotype of the indicated knock-out mice. IMPC: International Mouse Phenotyping Consortium (see: https://www.mousephenotype.org/data/genes/MGI:1341851). PMID: Pubmed ID. Source data are provided.
