Journal: The Journal of Biological Chemistry

Article Title: Selective Recognition of H3.1K36 Dimethylation/H4K16 Acetylation Facilitates the Regulation of All- trans -retinoic Acid (ATRA)-responsive Genes by Putative Chromatin Reader ZMYND8 *

doi: 10.1074/jbc.M115.679985

Figure Lengend Snippet: The list of antibodies used in Western blotting, co-IP, ChIP, and immunofluorescence

Article Snippet: Anti-GST-HRP , GE HealthCare , RPN1236.

Techniques: Western Blot

Journal: Journal of Biological Chemistry

Article Title: Phosphatidylinositol Phosphate Kinase Type Iγ Directly Associates with and Regulates Shp-1 Tyrosine Phosphatase

doi: 10.1074/jbc.m500576200

Figure Lengend Snippet: FIG. 1. PIPKI661 directly interacts with Shp-1. A, reciprocal GST pull-down of PIPKI661 and Shp-1. T7-tagged PIPKI661 was incubated with GST-Shp-1 or GST alone, and the complexes were pelleted with glutathione-Sepharose beads. Bound PIPKI661 was an- alyzed by Western blot using an antibody specific for the T7 epitope tag. T7-tagged Shp-1 was incubated with GST-PIPKI661, and the pelleted complex was analyzed in the same manner. The GST-tagged proteins were blotted with an antibody specific for GST. B, HEK293 cells were lysed, and Shp-1 was immunoprecipitated with a monoclonal antibody specific for Shp-1. The immunoprecipitates were resolved via SDS- PAGE and analyzed by Western blot as indicated. Normal mouse IgG was used as a control. C, HEK293 cells were co-transfected with 2 g of PIPKI661 and 2 g of Shp-1 via calcium phosphate. After 48 h, cells were subjected to immunoprecipitation assays using polyclonal antibod- ies specific for the C terminus of PIPKI661 or a monoclonal anti-Shp-1 antibody. The immunoprecipitates were resolved and probed with the antibodies indicated. Normal mouse and rabbit IgGs were used as controls.

Article Snippet: Horseradish peroxidase-conjugated anti-GST antibody was purchased from Amersham Biosciences, and horseradish peroxidase-conjugated anti-T7 an- tibody was obtained from Novagen.

Techniques: Incubation, Western Blot, Immunoprecipitation, SDS Page, Control, Transfection

Journal: Journal of Biological Chemistry

Article Title: Phosphatidylinositol Phosphate Kinase Type Iγ Directly Associates with and Regulates Shp-1 Tyrosine Phosphatase

doi: 10.1074/jbc.m500576200

Figure Lengend Snippet: FIG. 2. The phosphatase domain of Shp-1 binds to the C termi- nus of PIPKI. A, HEK293 cells were co-transfected with 2 g of either Shp-1 or Shp-2 and 2 g of the indicated PIPKI constructs. After 48 h, Shp-1 and Shp-2 were immunoprecipitated with monoclonal antibodies specific for the respective phosphatase. The immunoprecipitates were then analyzed by Western blot. The images blotting for immunoprecipi- tated PIPKI were taken from the same blot and exposure time. B, HEK293 cells were co-transfected as described in A with Shp-1 and the HA-tagged PIPKI constructs indicated (I483x, the codon for residue 483 of PIPKI661 was mutated into a stop codon, resulting in trunca- tion of the C terminus from that residue). The immunoprecipitates were then analyzed by Western blot as indicated. C, GST pull-down of PIPKI661 by Shp-1 truncations. T7-tagged PIPKI661 was pulled down by the GST-tagged Shp-1 constructs indicated. Bound PIPKI661 was detected by immunoblot with anti-T7 antibody. GST-tagged Shp-1 constructs were as follows: WT, wild type Shp-1; SH2, truncation of the Shp-1 C terminus from residue 278; PPase, truncation of the Shp-1 N terminus up to residue 300; GST, GST only. D, GST pull-down of Shp-1 C-terminal truncations. T7-tagged Shp-1 constructs were pulled down with GST-PIPKI661 and detected as described in C. C-terminal Shp-1 truncations: Shp-1 560x, C-terminal truncation of residues 560–597 by mutation of residue 560 into a stop codon; Shp-1 529x, C-terminal truncation of residues 529–597 in the same manner. Wild type T7- tagged Shp-2 (Shp-2) was used as an additional control.

Article Snippet: Horseradish peroxidase-conjugated anti-GST antibody was purchased from Amersham Biosciences, and horseradish peroxidase-conjugated anti-T7 an- tibody was obtained from Novagen.

Techniques: Transfection, Construct, Immunoprecipitation, Bioprocessing, Western Blot, Residue, Mutagenesis, Control