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Figure 6 | BMC Molecular Biology

Figure 6

From: Key role of the 3' untranslated region in the cell cycle regulated expression of the Leishmania infantum histone H2A genes: minor synergistic effect of the 5' untranslated region

Figure 6

Effect of the H2A4 5' UTR on regulation of CAT expression. (A) Protein extracts from 35S-labelled L. infantum promastigotes transfected with pXCAT5'H2A and treated with 5 mM HU either for 12 h (lane 0) or at the indicated time period (in h) after removal of the drug, were used to immunoprecipitate CAT. After immunoprecipitation, protein samples were separated by SDS/PAGE and transferred on to a nitrocellulose membrane. Autoradiography (De novo panel) or Western blot with anti-CAT antibodies to reveal the total amount of CAT present in the samples (TP panel) is shown. (B) Densitometric analysis showing the ratio between the de novo synthesis of CAT and total amount of CAT (columns). The percentage of cells in the S phase is also included (black line). Northern blots prepared with RNA samples separated by sucrose gradients from promastigotes transfected with pXCAT5'H2A (C) or pXCATHSP83 (D) treated 12 with HU (lane 0) or 3 h after drug removal were sequentially hybridized with the CAT and the H4 probe. The autoradiographic exposure of the blots and the densitometric analysis is shown. Data correspond to one representative experiment of three independent assays with similar results.

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