Fig 2: Differential expression analysis of MTDH and MYBL2 in normal tissues and glioma cancer tissues (a). Differential expression of two genes on the GEPIA online platform based on TCGA, the ordinate is the negative logarithmic transformation value of TPM+1 (b). In HEB, U251, U87, T98G, and LN-229 five cell lines. The relative expression levels of two genes in HEB, U251, U87, T98G, and LN-229 cell lines. N = 3, **p < 0.01 vs. the HEB group.

Fig 3: Protein-protein interaction networks of genes associated with neurogenesis and ion homeostasis identified by the Cytoscape software. (A) Changes in neurogenesis-associated genes after AEG-1 knockdown. A network of genes downregulated are shown. (B) Changes in the expression of genes associated with ion homeostasis, including eight genes that were upregulated and 22 genes that were downregulated.

Fig 4: The protein expression of MTDH, MYBL2, and FoxM1 in each group in U251 and U87 cell lines (a). The protein bands of the three genes in the five groups of control, NC, pc-MTDH, and siMTDH and the histogram of the relative protein expression; N = 3, **p < 0.01 vs. the NC group; ^^p < 0.01 vs. the siNC group (b). The three genes in siNC+pc-control, siMTDH+. The histogram of protein bands and relative protein expression in the three groups of pc-control and siMTDH+pc-FoxM1. N = 3, **p < 0.01 vs. the siNC+pc-control group; ^^p < 0.01 vs. the siMTDH+pc-control group.

Fig 5: Miat binds Mtdh to regulate microRNAs involved in stem cell maintenance, tumorigenesis, and treatment resistance. (A) RNA immunoprecipitation confirmed the association of Mtdh and Miat in stem-like MB cells. (B) Western blot of Mtdh and ß-actin in stem-like MB cells in two cell Mtdh-/- cell lines generated by CRISPR and Mtdh+/+ control. (C) Neurosphere formation in Mtdh-/- stem-like MB cells and Mtdh+/+ controls (n = 3). (D and E) Expression of genes associated with cancer stem cells (D) or stem cell differentiation (E) in Mtdh-/- stem-like MB cells and Mtdh+/+ controls (n = 3). (F) Cell viability as measured by MTS assay 48 h after increasing doses of IR (0, 1, 2, 3, 4, 6, 8, 10 Gy in Mtdh+/+ and Mtdh-/- stem-like MB cells; n = 4 in each group). (G) Heat map of differentially expressed small RNAs from small RNA-seq in stem-like MB cells generated with Cas9 and indicated sgRNAs targeting either Miat or nontargeting control Gal4-4 (Left) and Mtdh+/+ and Mtdh-/- stem-like MB cells (Right). (H) Correlation of magnitude and direction of change in 11 small RNAs that were significantly altered in sgMiat and Mtdh-/- stem-like MB cells compared to controls. Top, Venn diagram indicating the overlap in small RNAs altered by sgMiat or Mtdh loss; P < 10-6 by the Fisher exact test. (I) Expression of miRNAs in stem-like MB cells with indicated sgRNAs targeting either Miat with Cas9 or nontargeting control (Gal4-4) and Mtdh-/- stem-like MB cells. ***P value < 0.001; **P value < 0.01; *P value < 0.05 via Student t test.