Figure 5
Knockdown of lncRNA-ATB upregulates miR-200c expression in keloid fibroblasts.
(A,B) Pearson’s correlation analysis of relative expression levels of lncRNA-ATB and those of miR-200c determined using qRT-PCR in keloid (K) tissue and keloid fibroblasts (KF) compared to levels normal skin (NS) tissue and normal fibroblasts (NF), respectively; (C) quantitative RT-PCR analysis of endogenous miR-200c in KF with lncRNA-ATB knockdown using an lncRNA-ATB specific shRNA (shRNA-ATB) compared to a control shRNA (shRNA-control); (D) relative expression levels of lncRNA-ATB in KF overexpressing lncRNA-ATB (Lv-ATB), Lv-Luc was used as an control; (E) relative expression levels of miR-200c in KF transfected with an empty vector (PC3), a vector carrying wild-type lncRNA-ATB (PC3-ATB), a vector carrying mutated lncRNA-ATB [PC3-ATB-mut(miR-200c)], or a vector carrying another lncRNA induced by TGF-β and carrying none miR-200c binding site (PC3-508851); (F) relative expression levels of lncRNA-ATB in KF overexpressing miR-200c compared with those overexpressing a control miRNA (miR-control) is measured at 96 h; (G) schematic representation of the predicted binding site of miR-200c on lncRNA-ATB transcript. The nucleotides in red are the seed sequences of miR-200c; (H) luciferase activity in KF cotransfected with miR-200c and luciferase reporters containing nothing (vector), lncRNA-ATB (lncRNA-ATB-wt), or mutant transcript (lncRNA-ATB-mut). Data are presented as the relative ratio of firefly luciferase activity to renilla luciferase activity.
