Fig. 6.

LINC01050 directly binds to miR-7161-3p which targets SPZ1 by binding to its 3’UTR. a qRT-PCR analysis of miR-7161-3p expression in KATO III cells transduced with lentiviral control shRNA (sh-NC) or sh-LINC01050. ^*P < 0.05. b qRT-PCR analysis of miR-7161-3p expression in KATO III cells transduced with pLVX vector or pLVX-LINC01050. ^**P < 0.01. c Diagram of the luciferase reporter vectors containing the wild-type (WT) or mutant (MUT) LINC01050 sequences, with the highly conversed putative miR-7161-3p binding sites indicated. In the HEK293T cells, the miR-7161-3p mimic reduced the luciferase activity of the WT reporter relative to the negative control, but had little impact on the MUT reporter activity. ^**P < 0.01; ns, not significant. d-e Detection of LINC01050 and miR-7161-3p by qRT-PR in immunoprecipitated RNA after performing an anti-AGO2 RIP in KATO III cells. IgG was the negative control. ^** P < 0.01. f Enrichment of AGO2 protein in pull-down assay performed using LINC01050 or a negative control (NC) incubated with cell extracts. g qRT-PCR analysis of SPZ1 mRNA expression in KATO III cells transfected with the NC or miR-7161-3p mimics. ^**P < 0.01. h Western blot analysis of SPZ1 protein expression in KATO III cells transfected with the NC or miR-7161-3p mimics. *P < 0.05. i Diagram of the luciferase reporter vectors containing the WT or MUT sequence of the SPZ1 3’UTR, with the highly conversed putative miR-7161-3p binding sites indicated. Luciferase activity of pmirGLO vectors containing the WT or MUT SPZ1 3’UTR sequence after co-transfection into HEK293T cells with the NC or miR-7161-3p mimics. ^**P < 0.01; ns, not significant