Journal: Cancer cell

Article Title: Cooperative Epigenetic Remodeling by TET2 Loss and NRAS Mutation Drives Myeloid Transformation and MEK Inhibitor Sensitivity

doi: 10.1016/j.ccell.2017.11.012

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Slides were incubated with anti-pErk1/2 (Cell Signaling, cat# 4370, 1 μg/ml) and anti-Mac1 (abcam, cat#ab133357, 1 μg/ml) for 5 hr, followed by 60 min incubation with biotinylated goat anti-rabbit (Vector Labs, cat# PK6101, 5.75 μg/ml) at 1:200 dilution.

Techniques: Plasmid Preparation, Recombinant, Electron Microscopy, SYBR Green Assay, Selection, DNA Methylation Assay, TA Cloning, Sequencing, Cell Viability Assay, Quantitative RT-PCR, Methylation Sequencing, Software

Journal:

Article Title: Rabbit Cells Expressing Human CD4 and Human CCR5 Are Highly Permissive for Human Immunodeficiency Virus Type 1 Infection

doi:

Figure Lengend Snippet: Rabbit cells support Nef, Tat, and Rev function. (A) To estimate CD4 downregulation by Nef, a vector encoding Nef and a vector encoding CD8 were cotransfected into SIRC-CD4 cells. Surface protein expression was detected by staining with a MAb against CD4 or CD8 followed by FACS analysis. Cotransfection was performed because plasmids typically enter the same cells. By gating on CD8+ cells, mainly cells transfected with pNef were analyzed for CD4 expression. (B) Tat function was measured in cells cotransfected with a reporter construct (pLTR-CAT) and a vector encoding Tat protein. (C) Rev function was measured in cells cotransfected with a reporter construct (pDM128) and a vector encoding Rev protein. All analyses were performed in triplicate. OD(405), optical density at 405 nm.

Article Snippet: To generate rabbit cells expressing human CD4, parental SIRC cells (a rabbit fibroblast-like cell line derived from the cornea of a normal rabbit with no detectable reverse transcriptase activity; American Type Culture Collection [ATCC CCL-60], Rockville, Md.) were transfected by the calcium phosphate method with an expression vector (pcDNA3; Invitrogen, Carlsbad, Calif.) encoding human CD4 and a neomycin resistance gene (pCD4neo [ 25 ]) and were selected in culture medium with 400 μg of neomycin (Life Technologies, Grand Island, N.Y.) per ml.

Techniques: Plasmid Preparation, Expressing, Staining, Cotransfection, Transfection, Construct

Journal:

Article Title: Rabbit Cells Expressing Human CD4 and Human CCR5 Are Highly Permissive for Human Immunodeficiency Virus Type 1 Infection

doi:

Figure Lengend Snippet: CCR5-dependent induction of syncytia in SIRC-CD4/CCR5 cells infected by BaL. Hematoxylin-eosin staining was performed 2 days after infection.

Article Snippet: To generate rabbit cells expressing human CD4, parental SIRC cells (a rabbit fibroblast-like cell line derived from the cornea of a normal rabbit with no detectable reverse transcriptase activity; American Type Culture Collection [ATCC CCL-60], Rockville, Md.) were transfected by the calcium phosphate method with an expression vector (pcDNA3; Invitrogen, Carlsbad, Calif.) encoding human CD4 and a neomycin resistance gene (pCD4neo [ 25 ]) and were selected in culture medium with 400 μg of neomycin (Life Technologies, Grand Island, N.Y.) per ml.

Techniques: Infection, Staining

Journal:

Article Title: Rabbit Cells Expressing Human CD4 and Human CCR5 Are Highly Permissive for Human Immunodeficiency Virus Type 1 Infection

doi:

Figure Lengend Snippet: Intracellular viral gene expression indicates vigorous viral replication. After fixation and permeabilization, SIRC cells infected with BaL were stained with MAb against intracellular p24 (p24i) antigen.

Article Snippet: To generate rabbit cells expressing human CD4, parental SIRC cells (a rabbit fibroblast-like cell line derived from the cornea of a normal rabbit with no detectable reverse transcriptase activity; American Type Culture Collection [ATCC CCL-60], Rockville, Md.) were transfected by the calcium phosphate method with an expression vector (pcDNA3; Invitrogen, Carlsbad, Calif.) encoding human CD4 and a neomycin resistance gene (pCD4neo [ 25 ]) and were selected in culture medium with 400 μg of neomycin (Life Technologies, Grand Island, N.Y.) per ml.

Techniques: Gene Expression, Infection, Staining