Figure 2
A fragment, HMGA2-sh-3p20, is cleaved from HMGA2-sh by Drosha/DGCR8 complex and Dicer. (A) The diagram of the interaction between Drosha/DGCR8 and the hairpin structure. (B) Drosha (or DGCR8) RIP-PCR of HMGA2 mRNA in HepG2 cells. (C) Effect of si-Dicer on luciferase activities of HMGA2-sh-mediated pGL3-HMGA2 was measured by luciferase reporter gene assays in HepG2 cells. (D) The diagram shows the predicted miRNAs targeting pGL-HMGA2. (E) Effect of si-Dicer on the expression of miR-590, miR-410, miR-150, miR-132/212, miR-145 and miR-186 was examined by qRT-PCR in HepG2 cells. (F) The diagram shows the designed primers for fragments derived from HMGA2-sh. (G) The fragments were screened by RT-PCR using designed primers in HMGA2-sh-overexpressed 293T cells. NC, purified water; Mock, without plasmid DNA; Vector presents empty plasmid DNA. The full bands images are given as Supplementary Fig. S6. (H) The identified fragment, HMGA2-sh-20, was validated by qRT-PCR analysis in HepG2 cells transiently transfected with HMGA2-sh. Every experiment was repeated three times. Error bars represent s.d. (n = 3), **p < 0.01; ***p < 0.001, Student’s t test.
