Fig. 2

Co-localization of miR-10b and U6 snRNA in human GBM cells and tumors. a Fractionation of glioma cells indicates that miR-10b is distributed in both cytosolic and nuclear compartments. Representative Western blotting of cytoplasmic, nuclear soluble, and nuclear insoluble fractions of glioma LN229 cells for Hsp90 (cytosolic) and Lamin B (nuclear) markers (left). MiR-10b levels in cytoplasmic, nuclear soluble, and nuclear insoluble fractions in LN229 and U251 glioma lines by qRT-PCR (mean ± SD, n = 3) (right). b Representative FISH images of miR-10b (Alexa FluorTM 546, red) and U6 (Alexa FluorTM 488, green) in cultured LN229 and U251 glioma cells and non-glioma HEK-293 and SH-Sy-5y cell lines with a fluorescently labeled probe, and nuclei stained with DAPI. Arrows mark the colocalization of miR-10b with U6. Quantification of the colocalization between miR-10b and U6 is presented in right panels. The percentage of cells with the colocalized miR-10b and U6 signals and the number of colocalization spots per cell are shown. At least 80 cells were analyzed for each cell line. P values were calculated using one-way ANOVA. c Representative FISH images of miR-10b or miR-21 (red) and U6 (green) in patient-derived GBM tissues with the corresponding fluorescently labeled probes, and nuclei stained with DAPI. Arrows mark the colocalization of miR-10b with U6. ** P < 0.01; *** P < 0.001