Journal: mBio

Article Title: Barrier-to-Autointegration Factor 1 (BAF/BANF1) Promotes Association of the SETD1A Histone Methyltransferase with Herpes Simplex Virus Immediate-Early Gene Promoters

doi: 10.1128/mBio.00345-15

Figure Lengend Snippet: Effect of BAF on viral transcript levels. HFF cells were transfected with NT control or BAF-specific siRNA and infected with HSV-1 at an MOI of 10. (A) Knockdown efficiency was confirmed by quantitative RT-PCR. (B to D) Cells were harvested at the postinfection times indicated, total RNAs were prepared, and IE ( ICP4 [B] and ICP27 [C]) and E ( ICP8 [D]) gene mRNA levels were measured by quantitative RT-PCR. The mRNA levels were normalized to 18S rRNA. Results shown are means and standard deviations from three independent experiments.

Article Snippet: The membrane was then incubated with antibodies specific for HSV ICP8 (1:5,000, rabbit serum 3-83 [ ]), HSV ICP4 (1:4,000, mouse monoclonal antibody [MAb] 58S [ ]), ICP27 (1:5,000, MAb; Eastcoast Bio), gD (1:5,000, MAb; Eastcoast Bio), BAF (1:2,000, rabbit serum; Millipore), GFP (1:2,000, rabbit serum; Abcam), lamin B1 (1:10,000, rabbit serum; Abcam), GAPDH (1:10,000, MAb; Abcam), and FLAG M2 (1:2,000, MAb; Sigma).

Techniques: Transfection, Control, Infection, Knockdown, Quantitative RT-PCR

Journal: mBio

Article Title: Barrier-to-Autointegration Factor 1 (BAF/BANF1) Promotes Association of the SETD1A Histone Methyltransferase with Herpes Simplex Virus Immediate-Early Gene Promoters

doi: 10.1128/mBio.00345-15

Figure Lengend Snippet: Knockdown of BAF reduces the size and numbers of viral RCs, but heterochromatin is excluded from the RCs. HFF cells were transfected with NT control or BAF-specific siRNA and infected with HSV-1 at an MOI of 10. At 8 hpi, the cells were fixed and processed for indirect immunofluorescence assays with antibodies specific for BAF (red in top row), ICP8 (red in bottom row), and H3K9me3 (green). Arrows indicate ICP8-stained RCs.

Article Snippet: The membrane was then incubated with antibodies specific for HSV ICP8 (1:5,000, rabbit serum 3-83 [ ]), HSV ICP4 (1:4,000, mouse monoclonal antibody [MAb] 58S [ ]), ICP27 (1:5,000, MAb; Eastcoast Bio), gD (1:5,000, MAb; Eastcoast Bio), BAF (1:2,000, rabbit serum; Millipore), GFP (1:2,000, rabbit serum; Abcam), lamin B1 (1:10,000, rabbit serum; Abcam), GAPDH (1:10,000, MAb; Abcam), and FLAG M2 (1:2,000, MAb; Sigma).

Techniques: Knockdown, Transfection, Control, Infection, Immunofluorescence, Staining

Journal: mBio

Article Title: Barrier-to-Autointegration Factor 1 (BAF/BANF1) Promotes Association of the SETD1A Histone Methyltransferase with Herpes Simplex Virus Immediate-Early Gene Promoters

doi: 10.1128/mBio.00345-15

Figure Lengend Snippet: BAF localizes to viral RCs. (A) HFF cells were infected with HSV-1 at an MOI of 10, and an indirect immunofluorescence assay was performed with antibodies specific for BAF and ICP8 at 8 hpi. Images were deconvoluted by the inverse filter algorithm in the AxioVision 4.8 image acquisition software. (B) HeLa cell expressing FLAG-BAF were infected with HSV-1 DG1 at an MOI of 50, harvested at 4 hpi, and immunoprecipitated (IP) with antibodies specific for FLAG or ICP8. ICP4, ICP8, VP16-GFP, and FLAG-BAF were detected with antibodies specific for ICP4, ICP8, GFP, and FLAG.

Article Snippet: The membrane was then incubated with antibodies specific for HSV ICP8 (1:5,000, rabbit serum 3-83 [ ]), HSV ICP4 (1:4,000, mouse monoclonal antibody [MAb] 58S [ ]), ICP27 (1:5,000, MAb; Eastcoast Bio), gD (1:5,000, MAb; Eastcoast Bio), BAF (1:2,000, rabbit serum; Millipore), GFP (1:2,000, rabbit serum; Abcam), lamin B1 (1:10,000, rabbit serum; Abcam), GAPDH (1:10,000, MAb; Abcam), and FLAG M2 (1:2,000, MAb; Sigma).

Techniques: Infection, Immunofluorescence, Software, Expressing, Immunoprecipitation