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. 2011 Oct 18;40(3):1356–1365. doi: 10.1093/nar/gkr843

Figure 2.

Figure 2.

Molecular basis of the post-transcriptional regulation mediated by miR-96 on GPC3. (A) Schematic representation of the eGFP-GPC3 transgene with the 3′-UTR being targeted by miR-96. (B–D) HuH7 cells were transduced once with lentiviral particles expressing the transgene. After one week, the TCN was calculated using genomic DNA extracted from the eGFP-GPC3-expressing HuH7 cell population. Then cells were transfected with the indicated small RNAs. Three days later, the eGFP protein expression and mRNA amount were measured and data were analyzed following the FunREG experimental pipeline (9). (B) Global post-transcriptional regulation (ANOVA: P < 0.0001; n = 6). (C) mRNA stability (ANOVA: P < 0.0001; n = 6). (D) Translation efficiency (ANOVA: P = NS; n = 6). Panel (E): EGFP-GPC3-expressing HuH7 cells were first transfected with the indicated siRNA. Twenty-four hours later, cells were transfected with miR-96. Forty-eight hours later, eGFP protein expression was analyzed by FACS and data normalized to that of siLuc (ANOVA: P < 0.0001; n = 3). ***P < 0.001.