Journal: Molecular and Cellular Biology

Article Title: Exclusive Ubiquitination and Sumoylation on Overlapping Lysine Residues Mediate NF-κB Activation by the Human T-Cell Leukemia VirusTax Oncoprotein

doi: 10.1128/mcb.25.23.10391-10406.2005

Figure Lengend Snippet: FIG. 1. Differential intracellular partitioning of colocalized Tax and SUMO or Tax and ubiquitin molecules. 293T cells were transfected with vectors expressing (A) Tax alone or (B) Tax in combination with HA-Ub or HA–SUMO-1 and stained by immunofluorescence with (A) an anti-Tax MAb or (B) the anti-Tax MAb (IgG2a), an anti-HA rabbit polyclonal antibody, and a MAb (IgG1) recognizing either the Golgi matrix marker GM130 (upper panels) or the promyelocytic leukemia protein (lower panels). The cells were analyzed by laser scanning confocal microscopy, with

Article Snippet: Samples were washed and, when biotinylated goat anti-mouse IgG1 was used as the secondary antibody, cells were incubated for 20 min with Cy5-conjugated streptavidin (Jackson ImmunoResearch).

Techniques: Ubiquitin Proteomics, Transfection, Expressing, Staining, Marker, Confocal Microscopy

Journal: Molecular and Cellular Biology

Article Title: Exclusive Ubiquitination and Sumoylation on Overlapping Lysine Residues Mediate NF-κB Activation by the Human T-Cell Leukemia VirusTax Oncoprotein

doi: 10.1128/mcb.25.23.10391-10406.2005

Figure Lengend Snippet: FIG. 6. Tax colocalizes with IKK complexes in the cytoplasm and with free IKK in the nucleus. (A) 293T cells were cotransfected with the vector for expression of either Flag-IKK or Flag-IKK, alone or in combination with wild-type Tax or the Tax mutant K4-8R. The cells were submitted to triple-immunofluorescence staining with the anti-Tax MAb (IgG2a), an anti-Flag MAb (IgG1), and an anti-IKK rabbit polyclonal antibody. (B) 293T cells were transfected as described for panel A, except that HA-Ub was expressed instead of the IKK subunits and an anti-HA MAb (IgG1) was used during the triple-immunofluorescence staining. The images represent projections of Z-series confocal images. A fivefold enlargement of a field in the overlay images (Zoom) and diagrams of the intensity of the fluorescence for each of the staining along lines crossing cellular structures (S, cytoplasmic speckles; F, cytoplasmic fibers; NE, nuclear envelope) are shown.

Article Snippet: Samples were washed and, when biotinylated goat anti-mouse IgG1 was used as the secondary antibody, cells were incubated for 20 min with Cy5-conjugated streptavidin (Jackson ImmunoResearch).

Techniques: Plasmid Preparation, Expressing, Mutagenesis, Staining, Transfection

Journal: Cell reports

Article Title: Assessing kinetics and recruitment of DNA repair factors using high content screens

doi: 10.1016/j.celrep.2021.110176

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: For the experiment combining the three PI3K inhibitors, the following antibodies were used: mouse IgG2a anti-V5 tag (Abcam # ab27671) 1:1000 followed by anti-mouse IgG2a Alexa488 (Jackson ImmunoResearch #115–547-186) 1:200, rabbit anti-PAR (Enzo # ALX-210–890A-0100) followed by anti-rabbit IgG Alexa 647 (Invitrogen #A-21244) 1:250 and mouse IgG1 phospho-H2AX (Ser139) (Millipore # 05–636) 1:1000 followed by anti-mouse IgG1 Rhodamine Red (Jackson ImmunoResearch #115–297-185) 1:100.

Techniques: Recombinant, Transfection, Electron Microscopy, Bicinchoninic Acid Protein Assay, Control, Negative Control, esiRNA, Plasmid Preparation, Software, High Content Screening, Microscopy, Cell Culture, Imaging