Figure 4.

Casp12 is a novel direct target of miR-155. (A) Immunoblotting and statistical analyses of CASP12 in CCl₄-induced mice after administration of YD (mean ± SD, n = 6). ∗∗P < 0.01 vs. the oil group; ^##P < 0.01 vs. the CCl₄-induced group. (B) Immunoblotting and statistical analyses of CASP12 in LPS-induced RAW264.7 cells after administration of YD (mean ± SD, n = 3). ∗∗∗P < 0.001 vs. control; ^##P < 0.01, ^###P < 0.001 vs. LPS-induced group (without YD). (C) The results of RT-PCR analyses of Casp12 in the CCl₄-induced mice after administration of YD (mean ± SD, n = 6). ∗P < 0.05 vs. the oil group; ^#P < 0.05 vs. the CCl₄-induced group. (D) The results of RT-PCR analyses of Casp12 in the LPS-induced RAW264.7 cells after administration of YD (mean ± SD, n = 3). ∗∗∗P < 0.001 vs. control; ^##P < 0.01, ^###P < 0.001 vs. LPS-induced group (without YD). (E) Binding sites of Casp12 and miR-155 were predicted by TargetScan. The bold letters represent putative miR-155 target binding sites in the Casp12 3′ UTR. (F) Luciferase reporter analysis was utilized to confirm the binding between miR-155 and Casp12. The relative luciferase activity was normalized to Renilla activity. ∗∗P < 0.01, ∗∗∗P < 0.001 vs. miR-NC. (G) The protein expression levels of CASP12 in cells overexpressing miR-155 mimic or its control were analyzed by Western blotting and densitometric analyses. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ns: no significant differences.