Fig. 7: Effects of MAZ WT and its mutants on CCND1 expression, HCC cell proliferation and xenograft tumor growth. | Nature Communications

Fig. 7: Effects of MAZ WT and its mutants on CCND1 expression, HCC cell proliferation and xenograft tumor growth.

From: G-quadruplexes promote the motility in MAZ phase-separated condensates to activate CCND1 expression and contribute to hepatocarcinogenesis

Fig. 7

a Domain structures of MAZ WT and its phase separation and/or G4-binding deficient or FUS-IDR fusion mutants. b, c Droplet formation (b) and punctum formation (c) of mCherry-MAZ WT and mutants. In (b), representative fluorescence and DIC images of droplets (left) and their quantification (right) are presented. Data are mean ± s.e.m. (n = 6 fields/each). Scale bar: 20 μm. In (c), punctum quantification is shown at right. Data are mean ± s.e.m. (n = 30 cells/each). Zoom: 5× magnification. Scale bar: 2 μm. d Effects of MAZ WT and mutants on CCND1 promoter activity. Data are mean ± s.d. (n = 3 biologically independent experiments). e–g Effects of altered MAZ expression on cell viability and CCND1 expression. In (e), HL-7702 cells were infected by lentivirus expressing MAZ WT and mutants. In (f, g), HepG2 (f) and SMMC-7721 (g) cells harboring sh-MAZ-3’UTR to silence endogenous MAZ were infected by lentivirus expressing MAZ WT and mutants. Cell viability was determined by proliferation assays (top panel), and CCND1 mRNA levels were examined by RT-qPCR (bottom panel). Data are mean ± s.d. (n = 3 biologically independent experiments). h–k Effects of MAZ WT and mutants on mouse xenograft tumor growth. HepG2 cells with sh-MAZ-3’UTR-mediated endogenous MAZ knockdown and MAZ WT or mutant expression were grafted into nude mice. Tumor growth curves in about 3 weeks (h) and images of excised tumors (i) are presented. MAZ and CCND1 mRNA levels (j) and IF analyses of Ki-67 expression in xenograft tumors (k) are displayed. In (h), data are mean ± s.d. (n = 8 per group). In (j), data are mean ± s.d. (n = 3 biologically independent experiments). In (k), data are representative of three biologically independent experiments. l A schematic model of G4-mediated recruitment of MAZ to form MAZ/CCND1-G4 condensates that compartmentalize coactivators to activate CCND1 gene and subsequently promote hepatocarcinogenesis. In (b–h, j), unpaired two-tailed Student’s t-test was used for statistical analysis. Source data are provided as a Source Data file.

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