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Figure 2 | Molecular Cancer

Figure 2

From: Inhibition of succinate dehydrogenase dysregulates histone modification in mammalian cells

Figure 2

Succinate dehydrogenase gene inhibition dysregulates histone methylation. (A) HEK293 cells were transfected in duplicate with ON-TARGETplus SMART pool siRNAs targeting SDHD or with a non-targeting control (100 nM) for 72 h. One half of the sample was used to prepare RNA and the other half was processed for histone extraction. The silencing efficiency of SDHD was analysed by RT-PCR (top panel). Gel lanes were loaded with 5 μg histone extract (bottom panel), and immuno blotting carried out with the antibodies indicated. Histone H3 was used as a loading control. Graph shows densitometric analysis of two independent experiments. (B) Hep3B cells were transfected in duplicate with ON-TARGETplus SMART pool siRNAs targeting SDHB or with a non-targeting control (100 nM) for 72 and 96 h. One half of the sample was used to prepare whole cell extracts and the other half was processed for histone extraction. Gel lanes were loaded with 20 μg total cell extract (top panel) or 5 μg histone extract (bottom panel), and immuno blotting carried out with the antibodies indicated. Tubulin and histone H3 were analysed as loading controls for whole cell and histone extracts, respectively. Graph shows densitometric analysis of three independent experiments. (C) HEK293 cells were transfected as in (B) and processed for analysis after 96 h. Graph shows densitometric analysis of two independent experiments. (D) Immunoblot analysis of wild-type pU6, and SDHB-silenced D11 and D20 cell lines. Samples were processed as described above. Graph shows densitometric analysis of three independent experiments.

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