Figure 5 | Scientific Reports

Figure 5

From: Essential Roles of RNA-binding Protein HuR in Activation of Hepatic Stellate Cells Induced by Transforming Growth Factor-β1

Figure 5

TGF-β1 promotes HuR binding to SphK1 mRNA and prolongs half-life of SphK1 mRNA.

(a) LX-2 treated with or without TGF-β1 was collected. RIP experiment was performed as described in methods. SphK1 mRNA was measured by qRT-PCR and the PCR products were size-fractionated in a 2% agarose gel. LX-2 were transfected with SCR siRNA (b) or HuR siRNA (c) before TGF-β1 treatment for 24 hours, the half-life of SphK1 mRNA was determined by ActD pulse-chase experiments. LX-2 was pre-treated without (d) or with (e) LMB for 2 hours before TGF-β1 treatment and SphK1 mRNA half-life was examined. (f) Cells were transfected with pcDNA-flag-HuR plasmids and the half-life of SphK1 mRNA was tested. Data are presented as the means ± SEM derived from at least three independent experiments.

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