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Fig. 2 | Molecular Cancer

Fig. 2

From: YTHDF2 reduction fuels inflammation and vascular abnormalization in hepatocellular carcinoma

Fig. 2

YTHDF2 deficiency promotes tumor growth, vasculature remodeling and metastasis. (a) Proliferative activity of SMMC7721-shCtrl and SMMC7721-shYTHDF2 cells grown for 5 d as assessed by CCK8 (left), or grown for additional 24 h of normoxia (Nx) or hypoxia (Hx) and assessed by WST1 assay (right). n = 4–6 biological replicates. (b) Numbers of endothelial tubes formed by HUVEC cocultured with indicated SMMC7721 cells under Nx or Hx for 24 h. n = 3 biological replicates. (c, d) Indicated SMMC7721 (c) or MHCC97H (d) cells were injected subcutaneously into NPG mice and tumor volumes were measured at indicated dates. n = 5–6 mice. (e) H&E staining of lung tissues, showing metastasis nodules originated from MHCC97H cells. n = 5 mice. Scale bar, 100 μm. (f) Microvessel density (MVD) in SMMC7721-derived tumors, as assessed by CD31 staining and quantification of microvessel areas. n = 5 mice. Scale bar, 100 μm. (g) Vascular mimicry in MHCC97H-derived tumors, as quantified by numbers of PAS+CD31 channels lined by tumor cells. n = 5 mice. Scale bar, 100 μm. (h-j) Macropscopic appearances (h), tumor numbers (i) and maximum tumor sizes of livers from 8.5-month old Ythdf2F/F and Ythdf2LKO mice injected with DEN and CCL4. n = 7–9 mice. (j) H&E staining of metastasis nodules in Ythdf2F/F and Ythdf2LKO mouse lungs. n = 7–9 mice. Scale bar, 50 μm. (k) Immunofluorescence staining of CD31 and NG2, showing microvessels and pericyte coverage in Ythdf2F/F and Ythdf2LKO mouse livers. n = 7–9 mice. Scale bar, 50 μm. Error bars indicate means ± SEM *P < 0.05, **P < 0.01, ***P < 0.001, **** P < 0.0001. P-values were determined by two tailed t-test

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