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. 2019 Oct 3;10(10):746. doi: 10.1038/s41419-019-1983-5

Fig. 3. Up-regulation of NEAT1 causes neurotoxicity.

Fig. 3

a, b NSC-34 cells were transfected with 0.25 μg/well of the U6-sgRNA(MS2)_EF1a-MS2-P65-HSF1 vector (Vec) or the guide RNA sequence against NEAT1 promoter-containing U6-sgRNA(MS2)_EF1a-MS2-P65-HSF1 vector (NEAT1) in association with 0.75 μg/well of the pcDNA3 vector (Vec) or the pAC152-dual-dCas9VP64-sgExpression vector (dCas9-VP64) on 6-well plate. At 72 h after the transfection, the quantitative real-time PCR analysis of NEAT1 was performed (a). The cell viability was detected by WST-8 assay (b). The data are presented as means ± SD (N = 3). Statistical analysis was performed by one-way ANOVA followed by the Tukey’s multi comparisons test. c–e NSC-34 cells were infected with adenovirus encoding mouse NEAT1_1 or FLAG-PR100 at an MOI of 400 together with adenovirus encoding LacZ or Cre-recombinase at an MOI of 40. At 48 h after the infection, the quantitative real-time PCR analysis of NEAT1 was performed (c). The cell viability was detected by WST-8 assay (d). The cell lysates were subjected to dot blotting analysis using indicated antibodies (e). The data are presented as means ± SD (N = 3). Statistical analysis for data obtained from (c) NEAT1_2 and (d) was performed by one-way ANOVA followed by the Tukey’s multi comparisons test. Statistical analysis for data obtained from (c) NEAT1_1 & NEAT1_2 was conducted by the unpaired t-test