Fig. 5: Atoh8 regulates the acquisition of transformed features.
a, Top, experimental design. Bottom, picture of soft agar colonies, representative of three independent experiments. b, Colony quantification (n = 3 independent experiments). c, FACS analysis showing the emergence of Bcl11b-tdTomatolow/Thy1low (BLTL) cells during transformation in Control and Atoh8 KD backgrounds. d, Graph depicting the emergence of BLTL cells (n = 3 independent experiments). e, Histograms depicting Atoh8 transcript levels in patients. The data are presented as the log2[ratio of Atoh8 fragments per kilobase of transcript per million mapped reads (FPKMs)] between malignant and healthy tissues in paired samples. f, Schematic of the experimental design. Cells were split for at least ten passages (30 d) before subsequent analyses. g, Volcano plot showing differentially expressed genes in Control versus Atoh8 KD transformed line. Each dot corresponds to a transcript. Blue dots represent log2[FC] > 1 or <−1 and adjusted P value < 0.00001. Benjamini–Hochberg-adjusted P values of the comparisons were computed using the limma-voom workflow modified two-sided t-test. h, Western blot for E-cadherin, Snail, Vimentin, Twist1 and Gapdh in Control and Atoh8 KD transformed cell lines. i, PCA of the scRNA-Seq data. j, Single-cell pseudotime trajectories. k, Heatmap of the top temporally expressed genes based on the Atoh8 KD cell trajectory. l, Xenograft tumour volume over time (n = 6 independent mice per group). m, Survival plot of the mice from l (n = 6 independent mice). In b, d and l, the data are presented as means ± s.d. Statistical significance was determined by two-tailed Student’s t-test (b, d and l) or Kaplan–Meyer test (m).
