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Fig. 5 | Molecular Neurodegeneration

Fig. 5

From: Protective paraspeckle hyper-assembly downstream of TDP-43 loss of function in amyotrophic lateral sclerosis

Fig. 5

Loss of paraspeckles promotes apoptosis in cells with disturbed miRNA biogenesis and activated dsRNA response. a-c Disruption of paraspeckles in cells with downregulated TDP-43 or Drosha promotes apoptotic death in neuroblastoma cells. Efficiency of NEAT1_2 knockdown and levels of a proapototic protein CHOP mRNA were analysed by qRT-PCR (n = 3). * and #p ≤ 0.05 (Mann-Whitney U-test) (a and b). In c, representative images and quantification of cleaved caspase 3 (CC3) positive cells are shown. *p < 0.05, ** and ##p < 0.01 (one-way ANOVA). Scrambled siRNA or NEAT1 siRNA was co-transfected with an siRNA targeting TDP-43 or Drosha, and cells analysed 36 h post-transfection. * and # indicate statistically significant difference as compared to cells transfected with only scrambled siRNA or only NEAT1 siRNA, respectively. Scale bar, 100 μm. d-f Disruption of paraspeckles promotes apoptosis in dsRNA-stimulated cells. Cells were transfected with scrambled siRNA or NEAT1 siRNA and stimulated with poly(I:C) 36 h post-transfection. Induction of CHOP by poly(I:C) over time in normal cells (d) and CHOP mRNA levels in paraspeckle-deficient and paraspeckle-sufficient cells after 8 h of poly(I:C) stimulation (e) were analysed by qRT-PCR (n = 4). **p < 0.01 (Mann-Whitney U-test). In f, representative images and quantitation of CC3-positive cells in cultures transfected with scrambled siRNA or NEAT1 siRNA and treated with poly(I:C) for 24 h are shown. *p < 0.05 (Mann-Whitney U-test). Scale bar, 100 μm. g and h Expression of cytotoxicity-associated ISGs is potentiated by loss of paraspeckles in cells depleted of TDP-43 or Drosha (g) or stimulated by poly(I:C) (h). N = 3, *p < 0.05 (Mann-Whitney U-test)

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