Fig. 3

Knock-down of YTHDF2 inhibits tumor growth and metastasis in vivo. a-i Subcutaneous tumor model (BALB/c nude mice). a The tumor growth curve of xenografts was plotted in shNC and shYTHDF2 group (n = 5 each group) by measuring the tumor size (width² × length × 0.52) with vernier caliper each 4 days. 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 at 40 days by in vivo imaging system in two groups. The values of above groups 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 between two groups. g The EMT associated proteins and YTHDF2 protein extracted from anatomized tumor xenografts were analyzed by western blot assay. GAPDH was the internal reference. h Total RNA was extracted from the tumor xenografts, and m⁶A levels were determined by m⁶A RNA dot-blot assay. Methylene blue staining was loading control. i Representative IHC staining micrographs of Ki-67, YTHDF2 in tumor xenografts were conducted. Scale bar = 100 μm. j-l Metastatic model (BALB/c nude mice). j 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. k 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. l Representative H&E staining of metastatic organs (adrenal gland and 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