Fig 2: Isolation, culture, and identification of Schwann cells (SCs) and astrocytes (ASs). (A) Cultivation of SCs from sciatic nerve tissue. (a,a′) Numerous fibroblasts proliferated on day 10 of tissue culture, while few SCs survived. (b,b′) SCs reached a high confluence on day 30 of culture and were digested. (c,c′) The primary SCs were elongated in a spindle shape. The right graph is the magnification of the white dotted box of the left graph (a–c, scale bar = 200 μm; a′–c′, scale bar = 50 μm). (B) Characteristic markers (GFAP, Sox10, p75NTR) were highly expressed in SCs (red). The nucleus was dyed by Hoechst 33342 (blue) (n = 3, scale bar = 50 μm). (C) ASs derived from the neonatal rat brain cortex, presenting a branched, irregular morphology (left figure, scale bar = 200 μm; right figure, scale bar = 50 μm). (D) Characteristic markers (GFAP, S100β) were highly expressed in ASs (red), and the nuclei were stained by Hoechst 33342 (blue) (n = 3, scale bar = 50 μm).

Fig 3: Knocking down SOX10 antagonized the pro‐stemness phenotypes induced by miR‐637 inhibitor. (A,B) The stemness of indicated LN229 cells was examined by neurosphere formation assay, with representative images of formed neurospheres shown in (A) and the quantification results in (B). (C,D) The expression of CD133 on the surface of indicated cells was examined by flow cytometry in (C) and the quantification results are shown in (D). (E,F) The expression levels of stem‐cell markers Nestin, Oct4, Nanog, CD133 and CD44 were measured by Western blotting in indicated cells (E) and quantified as the ratio to GAPDH (F). Data are presented as mean ± SD from three independent experiments. Comparison between three or more groups was performed using one‐way ANOVA followed by Tukey’s post hoc test. *P < 0.05, **P < 0.01 and ***P < 0.001.

Fig 4: Critical role of miR-25-5p inhibitor in MLT+Ptero-treated colon cancer cells. (A) The binding sequences (red highlighted seed sequence) between 3′-UTR NEDD9 and hsa-miR-25-5p target genes by using miRWalk software. (B) Effect of miR-25-5p inhibitor on NEDD9 or SOX10 expression in MLT+Ptero-treated HT29 and SW480 cells. Expression levels of NEDD9 and SOX10 were analyzed by a Delta Vision Imaging System (Applied Precision). The nuclei were stained with DAPI (scale bar, 15 μm). (C) Effect of MLT+Ptero on NEDD9 in HT29 cells by qRT-PCR. After MLT+Ptero co-treatment for 24 h in HT29 cells, qRT-PCR was performed with total RNA isolated from HT29 cells. ** p < 0.01, *** p < 0.001 vs. miRNA inhibitor negative control. Data represent means ± SEM of triplicate samples.

Fig 5: SOX10 was a direct target for miR‐637 in glioma cells. (A–D) The expression levels of SOX10 were measured by qRT‐PCR (A,B) or by Western blotting (C,D) in LN229 or SHG‐44 cells treated with miR‐637 mimics or inhibitor. (E) Bioinformatic analysis identified a potential binding site between miR‐637 and the 3’‐UTR sequences of SOX10. (F) The luciferase activity was measured by reporter assay. Wild‐type (WT) or mutated (MUT) SOX10 binding sequence was cloned into the reporter construct, co‐transfected into LN229 or SHG‐44 cells with either miR‐637 mimics or inhibitor. Data are presented as mean ± SD from three independent experiments. Comparison between two groups was performed using Student’s t‐test and between three or more groups using one‐way ANOVA followed by Tukey’s post hoc test. *P < 0.05, **P < 0.01.