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. 2017 Feb 21;7:42882. doi: 10.1038/srep42882

Figure 7. hnRNP A1 contributes to rhythmic IRES-dependent translation of Nfil3.

Figure 7

(a) MC3T3-E1 cells were treated with 10 μM phenylephrine and transfected 8 hours with in vitro transcribed reporter mRNA (RF-Nfil3). Samples were harvested at the indicated times and subjected to a luciferase reporeter assay. Endogenous Nfil3 protein levels at indicated time points shown by immunoblotting. The ratio of FLUC/RLUC of cells transfected 12–20 hours after phenylephrine treatment was set to 1.0. Error bars represent mean ± SEM (n = 3), ***P < 0.001, **P < 0.01. Full-length blots are presented in Figure S11 and band of interest is indicated by a red box. (b) MC3T3-E1 cells were trated with phenylephrine and harvested at the indicated time points. Cells were lysed separately, and cytosolic and nuclear extracts were subjected to immunoblotting. Changes in cytosolic hnRNP A1 protein level were normalized to 14-3-3ζ protein. The ratio of hnRNP A1/14-3-3ζ protein at 16 hours after phenylephrine treatment was set to 1.0. This result is representative of three independent experiments. Full-length blots are presented in Figure S12 and band of interest is indicated by a red box.