Figure 6.

TGFB3-AS1 inhibits maturation of miR-144 to upregulate Rap1a expression in macrophages

(A) Localization of mature miR-144, pri-miR-144, and pre-miR-144 in macrophages was detected by qRT-PCR. (B) The stem-loop sequence of pri-miR-144 is partially complemented with TGFB3-AS1. (C) The levels of mature miR-144, pre-miR-144, and pri-miR-144 in macrophages after overexpression of TGFB3-AS1 (lenti-TGFB3-AS1) or knockdown of TGFB3-AS1 (TGFB3-AS1-sc). (D) The expression of Drosha was determined by qRT-PCR and western blot in macrophages after transduction with Drosha-expressing adenoviruses (Ad-Drosha). (E and F) The levels of mature miR-144, pre-miR-144, and pri-miR-144 were analyzed by qRT-PCR in macrophages co-transduced with Ad-Drosha and lenti-TGFB3-AS1 or TGFB3-AS1-siRNA in the presence of 100 μmol/L Hcy treatment. (G) Anchoring of DGCR8 to pri-miR-144 was determined by RIP assay in macrophages after TGFB3-AS1 was overexpressed. (H) The expression of Rap1a in the macrophages co-transduced with lenti-TGFB3-AS1 and miR-144 mimic in the presence of Hcy. (I) The expression of Rap1a in the macrophages after co-transduction with TGFB3-AS1-siRNA and miR-144 inhibitor in the presence of Hcy. ∗p < 0.05, ∗∗p < 0.01 compared with lenti-GFP or control group, #p < 0.05, ##p < 0.01 compared with TGFB-AS1-sc or 100 μmol/L Hcy, ▵p < 0.05, ▵▵p < 0.01 compared with 100 μmol/L Hcy + lenti-TGFB3-AS1 or 100 μmol/L Hcy + TGFB3-AS1-siRNA.