Fig. 3

Depletion of METTL16 inhibits CCA growth in mice. (A-C) CCLP1 cells transfected with CRISPR/Cas-9 vector carrying METTL16-specific guide RNA (sgMETTL16-2) or control guide RNA (sgCtrl) were subcutaneously inoculated into SCID mice. Tumor volumes measured at indicated time points are shown in (A). Gross photographs of tumors recovered from mice sacrificed at the end time point are shown in (B). Tumor weights from control vs METTL16-depleted group are shown in (C). D-F HuCCT1 cells transfected with CRISPR/Cas-9 vector carrying METTL16-specific guide RNA (sgMETTL16-2) or control guide RNA (sgCtrl) were subcutaneously inoculated into SCID mice. Tumor volumes (D), gross photos (E), and tumor weights F are shown. (G-J) CCLP1 cells transfected with CRISPR/Cas-9 vector carrying METTL16-specific guide RNA (sgMETTL16-2) or control guide RNA (sgCtrl) were inoculated into the livers of SCID mice. The mice were sacrificed 6 weeks post-inoculation. Gross photographs of livers recovered from each group are shown in (G). The liver/body weight ratios of mice from each group are shown in (H). The expression of the cell proliferation marker Ki67 and the apoptosis marker cleaved caspase-3 was determined by IHC analysis (I). CRISPR/Cas9-mediated depletion of METTL16 in the tumor tissues recovered from mouse livers was confirmed by Western blotting (J). **P < 0.01, ***P < 0.001, ****P < 0.0001. A, D Mean ± SD, One-way ANOVA test. C, F, H Unpaired t-test