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. 2013 Apr 19;41(11):5692–5703. doi: 10.1093/nar/gkt279

Figure 3.

Figure 3.

Subcellular localization of d2EGFP mRNA. HeLa Tet-Off®-derived cells were treated with either 0 or 1 µg/ml of doxycycline (shaded and open bars, respectively) to inhibit reporter gene expression and 4 h later these cells were collected. Total cellular, nuclear, cytoplasmic and polysomal RNA fractions were isolated. (A) A representative sucrose gradient sedimentation profile of the polysome preparation is provided for reference. Polysomal mRNAs are contained in fractions 3–5, as indicated. (B) The relative expression of d2EGFP and beta actin across these gradient fractions was determined by qRT-PCR. RNA levels are expressed relative to total input RNA level from no doxycycline control samples. (C) For comparison, the expression of d2EGFP mRNA following doxycycline treatment in (B) is expressed as a percentage of its respective total input RNA. (D) The expression of d2EGFP in several distinct cellular compartments was determined. Each value represents the mean (±SEM) from several independent experiments. The asterisk indicates the value is significantly different from controls in the presence of doxycycline, as determined by one-way ANOVA followed by a Tukey’s multiple comparisons test (P < 0.05).