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Figure 3 | Virology Journal

Figure 3

From: CHD1 and CHD2 are positive regulators of HIV-1 gene expression

Figure 3

CHD2 and CHD1 regulate retroviral infection in human cells. (A) siRNAs targeting CHD1 and CHD2 were transfected into HEK293T cells. A non-targeting siRNA (Neg) and a siRNA targeting the virus-encoded luciferase (GL3) were used as controls. (Left panel) Immunoblot analysis of CHD1, CHD2 and tubulin protein levels at 48 hours post-transfection. (Right panel) Protein levels were quantified using the Licor Odyssey infrared imaging system and the ratios of CHD1 or CHD2 protein:tubulin were determined and compared to the non-targeting control (defined as 100% Control). The data shown is the average mean of three independent experiments. (B) HEK293T cells transfected with the siRNAs were challenged with the MLV vector or (C) the HIV-1 vector. The ratios of luciferase activity:cell viability were determined and compared to the non-targeting control (defined as 100% infection). (D) siRNAs targeting both CHD1 and CHD2, or a control siRNA (Neg) were transfected into SupT1 cells. (Right panel) The cells were challenged with the HIV-1 vector as described above. (Left panel) CHD1 and CHD2 mRNA levels were measured by qRT-PCR. The ratios of CHD1 or CHD2 mRNA:GAPDH mRNA were determined and compared to the non-targeting control (defined as 100% Control). (Right panel) The ratio of virus-encoded luciferase activity:viable cell number was determined and compared to the non-targeting control (defined as 100% infection). (E) Immunoblot analysis of CHD1-V5, CHD2-Myc/His and tubulin protein levels at 48 hours post-transfection in transfected HEK293T cells. (F) The cDNA-expressing cells were challenged with the HIV-1 vector at 48 hours post-transfection. The ratio of virus-encoded luciferase activity:viable cell number was determined and compared to the non-targeting control (defined as 100% infection). The data shown is the average mean of three independent experiments each performed with triplicate samples. Error bars indicate the standard deviation. The data was analyzed using an unpaired T-test, ***P value <0.0001.

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