Fig. 5
Knock-down of METTL3 inhibits tumor growth and metastasis in vivo. a-h Subcutaneous tumor model (BALB/c nude mice). a The tumor growth curve of xenografts was plotted in shNC and shMETTL3 group (n = 5 each group) by measuring the tumor size (width2 × length × 0.52) with vernier caliper. The tumor size at the endpoint in above group was analyzed with student’s t test. b The subcutaneous tumor models were observed at 40 days in two different groups (blank arrows indicated tumor xenografts). c and d The luciferase activities (radiance values) of subcutaneous tumor xenografts were measured by in vivo imaging system in two groups. The values of above were analyzed with student’s t test. e The BALB/c nude mice were sacrificed for the xenografts, and the size was measured by the beside ruler. f The anatomized subcutaneous tumor xenografts were weighed and analyzed with student’s t test. g The EMT associated proteins and METTL3 protein extracted from anatomized tumor xenografts were analyzed by western blot assay. GAPDH was the internal reference. h Representative IHC staining micrographs of Ki-67, METTL3 in tumor xenografts were conducted. Scale bar = 100 μm. i-k Metastatic model (BALB/c nude mice). i The BALB/c nude mice injected with cells (1.5 × 106 per mouse) via tail vein were in vivo imaged at 4th weeks and 6th weeks to evaluate the whole metastasis. j The mice were sacrificed for the metastatic organs which were further in vivo imaged to reconfirm the metastasis. The representative photographs and corresponding gross specimens (right panel) were presented. k H&E staining of several metastatic organs (lung) were performed to identify the metastasis loci. Error bars represent the SD obtained from at least three independent experiments; *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001