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. 2013 Jun 25;41(16):7905–7919. doi: 10.1093/nar/gkt565

Figure 2.

Figure 2.

miR-195 represses Stim1 expression by destabilizing stim1 mRNA. (A) Levels of miR-195 in cells transfected with pre-miR-195 for 48 h as measured by Q-PCR analysis: (a) miR-195 levels; and (b) U6 RNA levels. Values are the means ± SEM from three separate experiments. *P < 0.05 compared with cells transfected with control scramble oligomer. (B) Changes in the levels of Stim1 and STIM2 proteins after ectopic miR-195 overexpression. Whole-cell lysates were prepared for western blotting; equal loading was monitored by assessing GAPDH levels. (C) Levels of stim1 mRNA as examined by Q-PCR (left) or RT-PCR (right) analyses. (D) Half-life of stim1 mRNA in cells overexpressing miR-195 as measured by Q-PCR analysis. Total cellular RNA was isolated at indicated times after administration of actinomycin D (5 µg/ml), and the remaining levels of stim1 and Gapdh mRNAs were measured by Q-PCR analysis. (E–H) Effect of miR-195 silencing on Stim1 expression. Forty-eight hours after cells were transfected with the corresponding oligomer targeting miR-195 (anti-miR-195) or control oligomer (C-oligo), various measurements were performed as described earlier in the text. Values are the means ± SEM from three separate experiments. *P < 0.01 compared with cells transfected with C-oligo.