Fig 2: Effect of the FAM83D-mediated AKT/mTOR pathway on the proliferation of glioma cells. A, MTT assays were conducted to evaluate the proliferation ability of U87 and U251 cells; B, Colony formation assays were conducted to determine the colony formation of U87 and U251 cells; C, Comparison of the colony numbers between U87 and U251 cells; * P < 0.05 vs. the Mock group; # P < 0.05 vs. the siFAM83D group; & P < 0.05 vs. the FAM83D group; % P < 0.05 vs. the MK2206 group.

Fig 3: Effect of the FAM83D-mediated AKT/mTOR pathway on the invasion and migration abilities of glioma cells. A, Transwell assays were conducted to determine the invasive abilities of U87 and U251 cells; B, Comparison of the invaded cell numbers between U87 cells and U251 cells; C, Wound healing tests were conducted to determine the migration abilities of U87 and U251 cells; B, Comparison of the wound closure between U87 cells and U251 cells; * P < 0.05 vs. the Mock group; # P < 0.05 vs. the siFAM83D group; & P < 0.05 vs. the FAM83D group; % P < 0.05 vs. the MK2206 group.

Fig 4: The effect of dietary methionine restriction on mTOR signaling and cancer progression. A Schematic of experimental design using HCC PDX model. Treatment, n = 5 mice per group. B Tumour growth curves and images of tumours at the end point (Day 28). C Survival analysis of mice under Ctrl or MR conditions. D Heatmap displaying global transcriptional changes of liver tissues between Ctrl and MR described in A. Red color represents the genes’ expression upregulated, while blue means the genes’ expression decreased. E KEGG pathway enriched for up-regulated and down regulated genes of liver tissues by comparing Ctrl to MR conditions. F Migration of SNU449 cells (left panel) with quantification data on the right panel. G Sphere formation efficiency of SNU449 cells (left panel) with quantification (right panel). H, I Tumour volumes and survival analysis of mouse allografts. n = 5 mice per group. Data are means ± SEM. Group differences were analyzed by two-tailed Student’s t test (B, C), two-way ANOVA followed by Tukey’s multiple comparison test (F, G, H) or log-rank test (I) (**p < 0.01, ****p < 0.0001). n.s., not significant. Ctrl, Control and MR, Methionine restriction

Fig 5: Expression of FAM83D and the AKT/mTOR pathway in U251 and U87 cells. A-B, Western blotting was conducted to determine the protein expression of FAM83D, p-AKT/AKT and p-mTOR/mTOR in glioma U251 (A) and U87 (B) cell lines; * P < 0.05 vs. the Mock group; # P < 0.05 vs. the siFAM83D group; & P < 0.05 vs. the FAM83D group; % P < 0.05 vs. the MK2206 group.