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. 2017 May 18;7:2070. doi: 10.1038/s41598-017-02311-0

Figure 3.

Figure 3

HMGA2-sh-3p20 upregulates HMGA2 by blocking the TTP-mediated degradation of HMGA2 mRNA. (A) The relative expression of HMGA2-sh-3p20 was assessed by qRT-PCR in 35 pairs of clinical HCC tissues and corresponding peritumor tissues (***P < 0.001; Wilcoxon’s signed-rank test). (B) The correlation between HMGA2 mRNA levels and HMGA2-sh-3p20 levels was measured by qRT-PCR in 30 cases of clinical HCC tissues (**P < 0.01, r = 0.586; Pearson’s correlation coefficient). (C) The luciferase activities of pGL3-HMGA2 were examined by luciferase reporter gene assays in HepG2 cells. (D) The expression of HMGA2 was assessed by qRT-PCR and Western blot analysis in Huh7 cells. (E) The diagram of TTP-mediated mRNA degradation. (F) The diagram of HMGA2-sh-3p20 antagonizes the interaction of TTP with non-hairpin within 3′UTR of HMGA2 mRNA. (G) TTP RIP-PCR of HMGA2 in HepG2 cells. (H) Effect of TTP on the expression of HMGA2 was measured by qRT-PCR and Western blot analysis in HepG2 cells. (I) Effect of HMGA2-sh-3p20 on the expression of TTP-mediated HMGA2 was assessed by qRT-PCR and Western blot analysis in HepG2 cells. The full length blots images are given as Supplementary Fig. S7. (J) TTP RIP-qPCR of HMGA2 in HepG2 cells transfected with HMGA2-sh-3p20. (K) Effect of HMGA2-sh-3p20 on the levels of HMGA2 mRNA in HepG2 cells transfected with si-TTP by qRT-PCR. (L,M) Effect of HMGA2-sh-3p20 on the half-life of HMGA2 mRNA in HepG2 cells (L) or HepG2 cells transfected with si-TTP (M) by qRT-PCR. Every experiment was repeated three times. Error bars represent s.d. (n = 3), **p < 0.01; ***p < 0.001 and not significant (NS), Student’s t test.