Fig. 3

miR-10b is enriched in spliceosomal SART3 and PRPF8 RNPs. a iCLIP of glioma cells for AGO2 and spliceosomal proteins LSM8, LSM4, SART3, PRPF8, DHX8, RBM22, and SNRNP200, followed by qRT-PCR for miR-10b and U6, demonstrates miR-10b enrichment in SART3 and PRPF8 RNPs. The results are expressed as fold-changes relative to the IgG (mean ± SD, n = 3). P values were calculated using two-tail unpaired t-test. b miR-10b-U6 chimeras were detected with a pair of PCR primers, one corresponding to miR-10b and another to U6, using iCLIP assay with AGO2 and Pan-Ago antibodies, and antibodies against LMNB and splieosomal factors LSM8, SART3, LSM1, LSM4, RBM22, PRPF8 in LN229 cells. c, d Representative images of miR-10b FISH (red) and either SART3 (c) or PRPF8 (d) immunofluorescence (green) in glioma cells, and nuclei stained with DAPI (blue) (n = 3). Arrows mark the miR-10b colocalization to SART3 and PRPF8 RNPs. e, f Representative images of the colocalization of miR-10b FISH (red) and either SART3 (e) or PRPF8 (f) immunofluorescence (green) in human GBM tumors, and nuclei stained with DAPI (blue) (n = 3). Arrows mark the miR-10b colocalization to SART3 and PRPF8 RNPs. g-j iCLIP with SART3 and PRPF8 antibodies on LN229 and GBM8 cells, transfected with either miR-10b inhibitor or mimic and the corresponding control oligonucleotides, followed by qRT-PCR detection of U6, demonstrate that U6 is displaced from the RNPs by miR-10b (mean ± SD, n = 3). P values were calculated using two-tail unpaired t-test. * P < 0.05; ** P < 0.01