Table 1.
Clinical information in infants with BA and CC.
Fig 1.
MiR-145 was predicted to target to ADD3 through 3’UTR interaction.
(A) and (B) ADD3 was predicted to have a putative miR-145 binding site within its 3’UTR.
Fig 2.
Luciferase reporter assays for miR-145 and ADD3 3’UTR co-transfection.
(A) MiR-145 repressed the luciferase activity by binding directly to ADD3 3’UTR in LX-2 cells, indicating a direct interaction between miR-145 and ADD3 in LX-2 cells. (B) NO differential luciferase activities were seen among miR-145, anti-miR-145 and NC groups, indicating miR-145 exactly binds to ADD3. UTR = untranslated region.
Fig 3.
Expression of ADD3in LX-2 cells in the intervention of lentiviral transfection.
(A) ADD3 mRNA was repressed byupregulated miR-145 transfection. (B) ADD3 protein expression was suppressed by miR-145.
Fig 4.
Evaluation of MiR-145 lentiviral transfection efficiency.
(A) Green fluorescence was detected in each group of lentiviral transfection except NC group, indicating a successful transfection. (B) qPCR was performed to quantify the expression level of miR-145-5p in each group, showing an increased miR-145-5p in both group 1 and 2 while not significantly decreased miR-145-5p in anti-miR-145 groups, indicating a low basic level of miR-145-5p in LX-2 cells.
Fig 5.
Pearson correlation analysis between miR-145-5p and ADD3.
The expression level of ADD3 was shown to be negatively reated to miR-145-5p in LX-2 cell. Pearson r was -0.9086 and 95% confidence interval was between -0.9901 and -0.3693, with an equation of Y = -0.8369*X + 1.496. ρ = 0.0121.
Fig 6.
Evaluation of the effect of miR-145 on Akt phosphorylation by Western blot analysis.
The protein levels of phosphorylated Akt (pAkt) was inhibited in both miR-145-5p and miR-145-transfected cells compared to NC groups.
Fig 7.
Expression of ADD3 at mRNA level in liver tissues.
ADD3 mRNA was quantified in 14 infants suffered from BA and 14 infants suffered from CC by qPCR. Compared with CC controls, the expression of ADD3 was significantly higher in BA group (p = 0.0118).
Fig 8.
Immunohistochemistry analysis for ADD3 expression in BA and CC liver biopsy tissues.
IHC for ADD3 was performed in BA and CC, showing a higher expression of ADD3 (red arrows) in BA group compared to CC group.
Fig 9.
MicroRNA gene microarrays in liver tissues from both BA and NC.
MiR-145 was shown to be differentially expressed in liver tissues of BA.
Fig 10.
Expression level of miR-145-5p in BA liver tissues.
qPCR was used to quantify the miR-145-5p expression in 14 infants with BA and 14 infants with CC. Compared with CC groups, the expression of miR-145 was significant in BA group (p = 0.0267)