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. 2015 Jul 7;4(7):e246. doi: 10.1038/mtna.2015.18

Figure 2.

Figure 2

8-mer LNAs against miR-372 and miR-373 de-repress LATS2 and inhibit the growth of cultured AGS cells. (a) Relative luciferase activity (mean ± SD, n = 3) of the LATS2 reporter system pGL3-LATS2wt containing the wt LATS2 3'UTR, or of the mutated LATS2 reporter system pGL3-LATS2mut, compared to that of the control vector pGL3. The vectors have been cotransfected with pRL-SV40 and 10 nmol/l of the indicated oligonucleotides using lipofectamin, and their expression was analyzed 48 hours later. (b) Western blot analysis of LATS2 (upper panel) and α-tubulin (lower panel) protein expression in untreated MKN74 cells (positive control) or in AGS cells treated with an equimolar mix of either 22-mer anti-miR-372+miR-373 antisenses (AS) or their scrambled sequence (SC), or with 8-mer LNA against miR-372 and miR-373 (TL) or their negative control (TLCo). (c) Relative growth rate (mean ± SD, n = 6) of tiny LNA-transfected cultured AGS cells between day 2 and day 5 post-treatment, compared to TLCo-treated cells. (d) LATS2 immunofluorescent staining in AGS cells in the same conditions than in (b); scale bars, 10 μm.