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. Author manuscript; available in PMC: 2015 Jan 12.
Published in final edited form as: Nat Commun. 2014 Nov 4;5:5248. doi: 10.1038/ncomms6248

Figure 3. AUF1 destabilizes NEAT1 and facilitates accumulation of exported mRNAs.

Figure 3

(a) RIP analysis was carried out to measure the relative enrichment of lncRNAs MALAT1 and NEAT1, as measured by RT-qPCR analysis (after normalization to GAPDH mRNA levels) in AUF1, HuR and TIA-1 RNPs or IgG. (b) Forty-eight hours after silencing AUF1, the half-life of NEAT1 RNA was measured as explained in the Methods section. (c) FISH analysis of NEAT1 and MALAT1 RNAs in HeLa cells 48 h after transfection of control, AUF1 or HuR siRNAs. DNA was counterstained with DAPI (blue). (d) Forty-eight hours after transfecting HeLa cells with control (Ctrl) or NEAT1 siRNA, RIP analysis was performed to measure the relative enrichment of PAICS, PCCB and NUP43 mRNAs (normalized to GAPDH mRNA) in AUF1 RNPs. (e) Relative distribution of PAICS, PCCB and NUP43 mRNAs in the nucleus and cytoplasm 48 h after transfection of control, AUF1 or HuR siRNA, respectively. (f) Levels of NEAT1 and MALAT1 RNAs (normalized to GAPDH mRNA) 48 h after silencing HuR or AUF1 in HeLa cells. Data in a,b,d–f are the means and s.d. from three independent experiments; *P<0.05, using Student’s t-test.