Extended Data Fig. 6: WM-8014 potentiates oncogene-induced senescence.
From: Inhibitors of histone acetyltransferases KAT6A/B induce senescence and arrest tumour growth
a, Growth curves of MEFs expressing empty vector control (pBABE) or oncogenic29 HRASG12V treated with increasing concentrations of WM-8014 as indicated or DMSO vehicle control. All experiments were performed in 3% O2. b, The effects of WM-8014 treatment in a zebrafish model of hepatocellular carcinoma19. Doxycycline-inducible, liver-specific expression of a GFP-krasG12V transgene leads to the accumulation of a constitutively active, GFP-tagged form of KRAS in hepatocytes. TO-krasG12V transgenic embryos were treated with doxycycline at 2 days post fertilization (dpf) and 5 dpf to initiate KRASG12V-driven hepatocyte proliferation. The size of the liver was measured by two-photon microscopy. Representative three-dimensional reconstructions of whole livers from image stacks after treatment of transgenic zebrafish Tg(TO-krasG12V) expressing KRASGV12 and GFP (green) in the liver or transgenic zebrafish Tg(lfabp10:RFP;elaA:eGFP) expressing only RFP (red). c, Quantification of liver volume. d, Incorporation of the nucleotide analogue 5-ethynyl-2′-deoxyuridine (EdU) after treatment of transgenic zebrafish expressing KRASG12V or control zebrafish with WM-8014 or control compound WM-2474. e, RT–qPCR determination of Cdkn2a (Ink4a) and Cdkn1a (encoding p21WAF1/CIP1) mRNA levels in transgenic zebrafish Tg(TO-krasG12V) treated as described in b. n = 6 independent cultures (a), 20 zebrafish (b, c), 10–12 zebrafish (d) and 4–5 zebrafish (e). Data are mean ± s.e.m. and were analysed by two-way ANOVA (a) or one-way ANOVA (d, e) followed by Bonferroni post hoc test with treatment and with or without treatment duration as the independent factors or by linear regression analysis regressing liver volume on WM-8014 concentration (c).
