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. 2013 May 22;8(5):e63745. doi: 10.1371/journal.pone.0063745

Figure 5. Spliced HIST1H2BD and HIST1H2AC transcripts are transported to the cytoplasm and found on polyribosomes.

Figure 5

(A) Polyadenylated HIST1H2BD and HIST1H2AC mRNA is transported to the cytoplasm. Cytoplasmic RNA was isolated from HCT116 cells treated either with DMSO or Nutlin-3a for 24 hours. RNA was reverse transcribed using random primers and analyzed for spliced HIST1H2BD and HIST1H2AC transcript by qRT-PCR. Values were normalized to 18S rRNA. Mean±SD, n = 3. (B) Representative polyribosome profiles obtained after sucrose gradient fractionation from DMSO and Nutlin-3a treated HCT116 cells. The x-axis represents the time of elution and y-axis represents the absorbance at 254 nm, indicating the RNA content. Polysome profiles were indicated in the figure. (C) RNA was extracted from the indicated polyribosome fractions of DMSO and Nutlin-3a treated cells and reverse transcribed using random primers. Expression of total and spliced HIST1H2BD and HIST1H2AC mRNA was analyzed by qRT-PCR and values were normalized to 18S rRNA. Mean±SD, n = 2. P-values were calculated and statistical significance is represented as follows (*P≤0.05; **P≤0.01).