Fig 1: miR-193b suppresses KRAS and STMN1 expression through their 3'-UTRs. (A) Schematic diagram of KRAS and STMN1 3'-UTRs with the locations of predicted conserved miRNA-targeting sequences highlighted. (B) Expression levels of KRAS and STMN1 were higher in F5M2 cells compared with F4 cells. (C) Luciferase reporter assay revealed that miR-193b suppressed luciferase activities of all WT constructs. (D and E) miR-193b negatively regulated the expression of KRAS and STMN1. (F) Immunofluorescence staining revealed that tumor tissue from nude mice injected with miR-193b-upregulated F5M2 cells exhibited lower expression levels of KRAS and STMN1 compared with the control group (magnification ×20). Data are presented as the mean ± SD of at least three independent experiments. *P<0.05. LV, lentivirus; miR-193b, microRNA-193b; MUT, mutant; NC, negative control; STMN1, stathmin 1; UTR, untranslated region; WT, wild-type.
Fig 2: miR-193b inhibits proliferation via KRAS and suppresses migration and invasion via STMN1. (A) Transfection efficiency was verified after F4 and F5M2 cells were transfected with siKRAS or pMSCV-KRAS, respectively. (B) Transfection efficiency was verified after F4 and F5M2 cells were transfected with siSTMN1 or pMSCV-STMN1, respectively. (C) Co-transfection with pMSCV-KRAS reversed miR-193b-mimic induced cell cycle arrest at G1 phase in F5M2 cells, whereas co-transfection with siKRAS reversed the miR-193b inhibitor-induced increase in percentage of cells at G2 phase in F4 cells. (D) Transwell cell invasion assay (magnification ×40). (E) wound-healing assay (magnification ×20) revealed that co-transfection of F5M2 cells with pMSCV-STMN1 and miR-193b mimic or co-transfection of F4 cells with siSTMN1 and miR-193b inhibitor reversed the regulatory effects of miR-193b mimic or inhibitor. Data are presented as the mean ± SD of at least three independent experiments. *P<0.05. miR-193b, microRNA-193b; NC, negative control; STMN1, stathmin 1; si, small interfering RNA.
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