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

Figure 6. hnRNP A1 regulates Nfil3 protein but not RNA oscillation.

Figure 6

(a) MC3T3-E1 cells were transfected with sh_M or sh_hnA1 expression vectors, incubated for 48 hours, treated with phenylephrine and harvested at the indicated time points. Samples were subjected to immunoblotting with the indicated antibodies. This result is representative of four independent experiments. Full-length blots are presented in Figure S10 and band of interest is indicated by a red box. (b) Relative sh_M (closed circles/solid line) and sh_hnA1 (open circles/dotted line) transfected Nfil3 protein were normalized to 14-3-3ζ protein, calculated and plotted. The ratio of sh_M transfected Nfil3/Gapdh protein at 12 hours after phenylephrine treatment was set to 1.0. Error bars represent mean ± SEM (n = 4). (c) MC3T3-E1 cells prepared in panel (a) were used for RNA quantification. Reverse-transcription and quantitative real-time PCR were performed using specific primers. Relative sh_M (closed circles/solid line) and sh_hnA1 (open circles/dotted line) transfected Nfil3 mRNA were normalized to Gapdh mRNA. The sh_M transfected Nfil3 mRNA level at 12 hours after phenylephrine treatment was set to 1.0. To statistically analyze the oscillation between sh_M and sh_hnA1, a two-way ANOVA was performed. Error bars represent mean ± SEM (n = 4). ns P ≥ 0.05.