Extended Data Fig. 4: Silencing RIOK1 inhibits pentose phosphate pathway and tumor growth. | Nature Cancer

Extended Data Fig. 4: Silencing RIOK1 inhibits pentose phosphate pathway and tumor growth.

From: RIOK1 phase separation restricts PTEN translation via stress granules activating tumor growth in hepatocellular carcinoma

Extended Data Fig. 4

a, KEGG pathway enrichment analysis of different metabolites in HCCLM3 cells with RIOK1 knockdown and ctrl group. b, Heatmap representing markedly changed metabolites associated with nucleotides and pentose phosphate pathway in HCCLM3 cells with RIOK1 knockdown and ctrl group, as determined using untargeted metabolomics (n = 5 independent cultures per cell line). c-d, Images depict the scheme to selectively inactivate the RIOK1 gene in murine livers by targeting exon 2 using Cre-lox technology (c). Representative RT–PCR-based genotyping assays to identify mice bearing the floxed RIOK1 allele (fl/fl) (top panel) in combination with albumin (Alb) driven expression of Cre recombinase (Alb-CreERT2 transgene) (bottom panel) (d). e-f, Construct design (e) and experimental scheme (f) for the spontaneous liver tumor model induced by hydrodynamic tail vein injection with the C-myc proto-oncogene transposon system in combination with CRISPR–Cas9 targeting of Trp53 (SgP53/Myc). g, Experimental scheme for the spontaneous liver tumor model induced by DEN/CCl4 treatment. h, Representative Ki67 stained liver sections were shown refer to Fig. 5g. n = 3 mice. i, Western blotting showing the expression of RIOK1, PTEN and G6PD in liver tumors from Fig. 5g; n = 6 mice/group). j, GO functional analysis diagram of markedly differentially expressed genes in RIOK1 knockout tumors (n = 5 mice/group). k-l, Hepatic spontaneous neoplasms were challenged by hydrodynamic tail vein (HTV) injection of Sleeping Beauty (SB) transposase vector encoding MycOE, Trp53KO in both RIOK1fl/fl and RIOK1ΔAlb group mice. In a separate cohort of mice, we administered a combination of SB/MycOE, IGF2BP1 Y396A, and Trp53KO plasmids. Representative freshly excised livers (k) and Liver/body weight ratios (l) indicative of tumor burden in the different genotypes (n = 3 mice/group). Data are represented as mean ± SD and analyzed by one-way ANOVA with Tukey’s multiple comparisons test.

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