Research Paper Volume 11, Issue 21 pp 9569—9580

TGF-β- and lipopolysaccharide-induced upregulation of circular RNA PWWP2A promotes hepatic fibrosis via sponging miR-203 and miR-223

Figure 4. Circ-PWWP2A regulates Fstl1 and TLR4 via sponging miR-203 and miR-223, respectively. (A) The luciferase reporter vector carrying mutant Fstl1 3’-UTR was transfected into 293T cells along with miR-203 inhibitor, and the luciferase activity, mRNA and protein expressions of Fstl1 were detected. (B) LX-2 cells were transfected with Ad-circ-PWWP2A or co-transfected with miR-203 mimic. The mRNA and protein expressions of Fstl1 were detected. (C) LX-2 cells were transfected with si-circ-PWWP2A or co-transfected with miR-203 inhibitor followed by TGF-β- or LPS-activation. The protein expression of Fstl1 was detected. (D) The luciferase reporter vector carrying mutant TLR4 3’-UTR was transfected into 293T cells along with miR-223 inhibitor, and the luciferase activity, mRNA and protein expressions of TLR4 were detected. (E) LX-2 cells were transfected with Ad-circ-PWWP2A or co-transfected with miR-223 mimic. The mRNA and protein expressions of TLR4 were detected. (F) LX-2 cells were transfected with si-circ-PWWP2A or co-transfected with miR-223 inhibitor followed by TGF-β- or LPS-activation. The protein expression of TLR4 was detected. **p<0.01 vs NC or Ad-GFP; ##p<0.01 vs Ad-circ-PWWP2A+mimic NC. Ad-GFP, adenoviral vectors carrying green fluorescent protein genes that act as the control. NC, negative control. si-circ-PWWP2A, small interfering RNA of circ-PWWP2A.