Fig. 3

Enhanced paraspeckle assembly in cells with compromised function of the miRNA pathway. a Downregulation of Drosha, Dicer, Ago2, ADAR1 and FUS in neuroblastoma cells after transfection of specific siRNA as analysed by qRT-PCR (n = 4–6). *p < 0.05, **p < 0.01 (Mann-Whitney U-test). b and c Knockdown of Drosha, Dicer, Ago2 or ADAR1 results in increased paraspeckle formation. Representative images of cells (b) and paraspeckle quantification (c) are shown. In c, the mean number of paraspeckles per cell and frequencies of such cells were plotted; the number of cells analysed is indicated at the bottom of each bar. *p < 0.05, **p < 0.01; ****p < 0.0001 (one-way ANOVA with Holm-Sidak correction for multiple comparisons). d NEAT1 is upregulated in cells after knockdown of Drosha, Dicer, Ago2 and ADAR1 (n = 4–6). *p < 0.05, **p < 0.01, ***p < 0.001 (one-way ANOVA with Holm-Sidak correction for multiple comparisons). e Suramin stimulates NEAT1_2 expression and paraspeckle formation. Cells were treated with suramin for 24 h before collection for RNA-FISH and qRT-PCR analysis (n = 4). *p < 0.05 (Mann-Whitney U-test). In a-d, cells were analysed 48 post-transfection. Scale bars are 10 μm in all panels