Figure 1.

NEAT1 expression and paraspeckle formation are induced by SFN. A, MCF7 cells were treated with SFN (20 μm) for the indicated time points. RNA was isolated, and the expression of NEAT1 (both isoforms) and NEAT1_2 was determined by RT-qPCR. The mean value ± S.D. of three biological replicates in one experiment is presented as fold change relative to untreated cells. The results are representative of three independent experiments. B, MCF7 cells were pre-incubated with NAC (5 mm) and then treated with SFN for 6 h. NEAT1 expression was determined as described in A. C, MCF7 cells were left untreated or treated with SFN for 6 h, fixed, and subjected to RNA-FISH using probes recognizing the NEAT1_2 isoform. DAPI was used to visualize the nuclei. Bars, 10 μm. D, overall intensity of the dots in at least 250 cells was quantitated using the Volocity software. Mean values ± S.D. of three biological replicates are shown and presented as fold change relative to untreated cells. p values were calculated using ANOVA (A) or Student's t test (B and D) with p < 0.05 considered statistically significant. (***, p ≤ 0.001; **, p ≤ 0.01.)