Skip to main content
. 2022 Feb 17;113(4):1154–1167. doi: 10.1111/cas.15272

FIGURE 6.

FIGURE 6

AUF1‐enhanced AKR1B10 expression through post‐transcriptional regulation. A, B, Western blot and RT‐qPCR analyses of AKR1B10 protein (A) and mRNA (B) in hepatoma cells with AUF1 overexpression or knockdown. C, Expression levels of AUF1 protein were positively correlated with AKR1B10 in 40 pairs of human HCC samples determined by western blot. Data were digitalized using ImageJ software statistical analysis. D, mRNA expression levels of AKR1B10 in 40 pairs of HCC samples were positively correlated with protein expression of AUF1. E, Half‐life of AKR1B10 mRNA in HepG2 and Huh‐1 cells with AUF1 knocked down. The levels of AKR1B10 mRNA was tested using qRT‐PCR at the indicated time followed by actinomycin D treatment (5 mg/mL). F, Binding of AUF1 to AKR1B10 mRNA detected using ribonucleoprotein immunoprecipitation (RNP‐IP) assays. AKR1B10 mRNA abundance in immune precipitates was tested using RT‐PCR and qRT‐PCR. G, Motif in AKR1B10 mRNA interacted with AUF1 identified using RNA pulldown assay. Ctrl indicates the negative control, and Input indicates total RNA extracted. 5′UTR, CR, and 3′UTR, respectively, indicate the corresponding regions of AUF1 mRNA. H, Luciferase activity of the reporter containing the 3′UTR or 5′UTR of the AKR1B10 transcript detected using luciferase activity assays. *P < .05; ***P < .001