DOT1L acetylation regulates CRC migration, invasion and metastasis in vivo. (A, B) Transwell cell migration (A) and matrigel cell invasion (B) assays in HCT116 cells transfected with either pcDNA, DOT1L(WT), DOT1L(K358Q) or DOT1L(K358R) plasmids (upper). The data represent the means ± SEM (n = 3) (lower). *p < 0.05, ***p < 0.001. (C) Comparative growth assay for pcDNA, DOT1L(WT), DOT1L(K358Q) and DOT1L(K358R) overexpressing HCT116 cells. The cell number of each sample was counted at the indicated times. (D, E) HCT116 cells stably expressing pHBLV-luci control (pC), pHBLV-luci-DOT1L(WT) pHBLV-luci-DOT1L(K358Q) or pHBLV-luci-DOT1L(K358R) plasmids were injected intravenously via the tail vein into 6-week-old male nude mice (n = 6 mice per group). Lung metastasis was monitored by bioluminescent imaging after 7 weeks of injection. Representative in vivo bioluminescent images and the incidence of lung metastasis from the different groups are shown (D); the bioluminescent quantitation of lung metastases is given (E). *p < 0.05 compared with the DOT1L(WT) and DOT1L(K358Q). The data represent the means ± SD. (F) Representative lung metastasis specimens were sectioned and stained with H&E. Scale bars: 200 μm.