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. 2016 May 13;6:25901. doi: 10.1038/srep25901

Figure 4. Knock down of LSD1 represses HBV transcription and replication.

Figure 4

(A) The scheme depicts the experimental procedure. Human hepatocarcinoma Huh7 cells were transfected with 10 nM of either control or LSD1 siRNA. 24 h after, cells were transfected with the HBV genome. (B) LSD1 knock down was determined by Western blot using specific antibodies. Beta actin was used as a loading control. (C–G) Covalent post-translational modifications on histone H3 were determined by ChIP analysis using the specific antibodies as indicated. Immunoprecipitated DNA was quantified by qPCR using specific primers as detailed in Material and Methods. When analyzing histone modifications, we normalized the data against the immunoprecipitated H3. The standard deviation was obtained from three PCR reactions and the graphs are representative of three independent experiments. (H) HBV cccDNA and cytoplasmic intermediate levels were determined by qPCR using specific primers. The results are shown as fold changes with respect to the control. The standard deviation was obtained from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, Student´s t-test.