Fig 1: miR-30d directly targets GNA13 3′-UTR. (A and B) Predicted targeting sites of miR-30d within 3′-UTR regions of GNA13. miR-30d targeting sequences are evolutionarily conserved in the 3′-UTR regions of GNA13 in 16 species. The targeting sites are highlighted in red. (C and D) Following the transfection of miR-30d mimic or NC mimic in SW480 cells, mRNA and protein expression of GNA13 was determined by (C) western blot analysis and (D) RT-qPCR *P<0.05 vs. the NC mimic (E) Five colon cancer cell lines were used to determine the expression levels of GNA13 mRNA and miR-30d using RT-qPCR. The trend of opposite expression was identified between expression levels of GNA13 mRNA and miR-30d in colon cancer tissues. β-actin and U6 were used as the endogenous controls, respectively. *P<0.05 vs. HCT116. (F) WT or MUT reporters and the miR-30d mimic or NC mimic were used to co-transfect 293T cells. Luciferase/Renilla activity was measured after 48 h. *P<0.05 vs. the WT NC mimic. miR, microRNA; NC, negative control; UTR, untranslated region; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; WT, wild-type; MUT, mutant.
Fig 2: Schematic diagram of G protein-coupled receptors and the underlying GNA13 mechanism leading to carcinogenesis and metastasis. LPA, lysophosphatidic acid.
Fig 3: Tumor suppressive effects of miR-30d via GNA13 expression inhibition. (A and B) Expression of (A) miR-30d expression and (B) GNA13 was evaluated by RT-qPCR and western blot analysis, respectively, in SW480 cells co-transfected with miR-30d mimic (or NC mimic) and GNA13 [or pcDNA3.1(+)]. *P<0.05 GNA13 vs. pcDNA3. (C-E) Transwell migration and invasion assays. (C) Representative images (magnification, ×200) of Transwell assays; (D and E) quantitative analysis of migration and invasion. *P<0.05, **P<0.01. (F) SW480 cells were transfected with siGNA13 or siNC. GNA13 expression was determined by western blotting after 48 h. (G and H) Following transfection with siRNA or siNC, Transwell migration and invasion assays were performed. (G) Representative images (magnification, ×200) of Transwell assays; (H) quantitative analysis of migration and invasion. *P<0.05, **P<0.01 vs. siNC. miR, microRNA; NC, negative control; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; siRNA or si, small interfering RNA.
Supplier Page from MyBioSource.com for GNA13 siRNA (Human)