Journal:

Article Title: AMP-Activated Protein Kinase Deficiency Exacerbates Aging-Induced Myocardial Contractile Dysfunction

doi: 10.1111/j.1474-9726.2010.00586.x

Figure Lengend Snippet: Western blot analysis depicting protein expression of ryanodine receptor (RyR), L-type Ca2+ channel dihydropyridine receptor (DHPR), sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA2a), Na+/Ca2+ exchanger (NCX), phospholamban (PLB) and phosphorylated PLB (pPLB, Ser16) in myocardium from young or old wild-type (WT) and AMPK deficient KD mice. (A) Representative gel blots of RyR, DHPR, SERCA2a, NCX, PLB, pPLB and GAPDH (loading control) using specific antibodies; (B) RyR expression; (C) DHPR expression; (D) SERCA2a expression; (E) NCX expression; (F) PLB expression; (G) pPLB (Ser16) expression and (H) pPLB-to-PLB ratio. Mean ± SEM, n = 3 – 6 mice per group, *p < 0.05 vs. respective young group, #p < 0.05 vs. WT-old group.

Article Snippet: Membranes were probed with anti-Akt (1:1,000), anti-pAkt (Thr 473 , 1:1000), anti-AMPK (1:1,000), anti-pACC (Ser 79 , 1:1,000), anti-HSP90 (1:1,000 ), anti-peNOS (Ser 1177 , 1:1,000), anti-Glut4 (1:1,000, all from Cell Signaling), anti-PGC-1α (1:500) anti-DHPR (1:200), anti-RyR (1:100, all from Santa Cruz Biotechnology, Santa Cruz, CA), anti-SERCA2a (1:1,000, Bethyl Laboratories Inc., Montgomery, TX), anti-PLB (1:1,000), anti-pPLB (Ser 16 , 1:1000 dilution; both from Upstate, Lake Placid, NY), anti- Na + /Ca 2+ exchanger (Abcam Inc., Cambridge, MA) and anti-GAPDH (1:1,000, internal loading control, Cell Signaling) antibodies.

Techniques: Western Blot, Expressing

Journal:

Article Title: EBP2, a Human Protein That Interacts with Sequences of the Epstein-Barr Virus Nuclear Antigen 1 Important for Plasmid Maintenance

doi:

Figure Lengend Snippet: Activation of the HIS3 reporter by EBNA1 and EBP2 in the two-hybrid system. Tenfold serial dilutions of log-phase cultures of Y190 strains expressing SNF1, EBNA1 or lamin as GAL4 DNA binding domain fusions and SNF4, EBP2, or nothing (pACT2) fused to the GAL4 activation domain were plated on SC-Trp,Leu (left panel) or SC-Trp,Leu,His plus 50 mM AT (right panel). The SNF1/SNF4 culture is a positive control for the two-hybrid interaction.

Article Snippet: A baculovirus expressing the tagged EBP2 was generated by homologous recombination of pVLEBP2 with Baculogold baculovirus DNA (Pharmingen, San Diego, Calif.) as previously described ( 4 ).

Techniques: Activation Assay, Expressing, Binding Assay, Positive Control

Journal:

Article Title: EBP2, a Human Protein That Interacts with Sequences of the Epstein-Barr Virus Nuclear Antigen 1 Important for Plasmid Maintenance

doi:

Figure Lengend Snippet: Interaction of EBNA1 and EBP2 in the one-hybrid system. Triplicate cultures of LF100 expressing EBNA1 (from pEBNA1) and EBP2 fused to the GAL4 activation domain (from pACT63) were grown in liquid SC-Trp,Leu,His plus 5 mM AT, and the growth was monitored by measuring the OD600 (triangles). The growth of negative control cultures containing pEBNA1 with pACT2 (circles) or pACT63 with pAS2 (squares) is also shown.

Article Snippet: A baculovirus expressing the tagged EBP2 was generated by homologous recombination of pVLEBP2 with Baculogold baculovirus DNA (Pharmingen, San Diego, Calif.) as previously described ( 4 ).

Techniques: Expressing, Activation Assay, Negative Control

Journal:

Article Title: EBP2, a Human Protein That Interacts with Sequences of the Epstein-Barr Virus Nuclear Antigen 1 Important for Plasmid Maintenance

doi:

Figure Lengend Snippet: The EBP2 cDNA. The cDNA isolated from the 5′-Stretch Plus library is shown. The Kozak consensus sequence is in boldface and the polyadenylation sequence is underlined.

Article Snippet: A baculovirus expressing the tagged EBP2 was generated by homologous recombination of pVLEBP2 with Baculogold baculovirus DNA (Pharmingen, San Diego, Calif.) as previously described ( 4 ).

Techniques: Isolation, Sequencing

Journal:

Article Title: EBP2, a Human Protein That Interacts with Sequences of the Epstein-Barr Virus Nuclear Antigen 1 Important for Plasmid Maintenance

doi:

Figure Lengend Snippet: Alignment of human EBP2 with the S. cerevisiae (YKL172w), Schizosaccharomyces pombe (SPAC17H9), and C. elegans (C18A3.3) homologues. The alignment was performed by using the Clustal method in DNASTAR. Identical and highly conserved amino acids are shaded.

Article Snippet: A baculovirus expressing the tagged EBP2 was generated by homologous recombination of pVLEBP2 with Baculogold baculovirus DNA (Pharmingen, San Diego, Calif.) as previously described ( 4 ).

Techniques:

Journal:

Article Title: EBP2, a Human Protein That Interacts with Sequences of the Epstein-Barr Virus Nuclear Antigen 1 Important for Plasmid Maintenance

doi:

Figure Lengend Snippet: Predicted structure of human EBP2. The predicted secondary structure (2°) of EBP2 was determined by using PHDsec (EMBL, Heidelberg, Germany). L, loop; H, α-helix (no β-sheets were predicted). Predicted structures are only shown for residues with secondary structure reliability indices of 5 or more. The position of coiled coils (C-C) was determined by using PairCoil, and residues with a 40% or greater probability of forming coils are shown (C).

Article Snippet: A baculovirus expressing the tagged EBP2 was generated by homologous recombination of pVLEBP2 with Baculogold baculovirus DNA (Pharmingen, San Diego, Calif.) as previously described ( 4 ).

Techniques:

Journal:

Article Title: EBP2, a Human Protein That Interacts with Sequences of the Epstein-Barr Virus Nuclear Antigen 1 Important for Plasmid Maintenance

doi:

Figure Lengend Snippet: Coimmunoprecipitation of EBNA1 and EBP2. Insect cells were infected with baculoviruses expressing EBNA1 or EBP2 alone or were coinfected with both baculoviruses. Lysates from metabolically labeled cells were prepared 3 days (A and C) or 2 days (B) postinfection and immunoprecipitated with anti-EBNA1 antibody (IP). (C) Immunoprecipitates from lysates coexpressing EBNA1 and EBP2 are shown before and after treatment with RNase A and DNase I.

Article Snippet: A baculovirus expressing the tagged EBP2 was generated by homologous recombination of pVLEBP2 with Baculogold baculovirus DNA (Pharmingen, San Diego, Calif.) as previously described ( 4 ).

Techniques: Infection, Expressing, Metabolic Labelling, Labeling, Immunoprecipitation

Journal:

Article Title: EBP2, a Human Protein That Interacts with Sequences of the Epstein-Barr Virus Nuclear Antigen 1 Important for Plasmid Maintenance

doi:

Figure Lengend Snippet: Coimmunoprecipitation of EBP2 with EBNA1 mutants. (A) Lysates from insect cells expressing EBNA1 proteins alone (−) or EBNA1 proteins with EBP2 (+) were prepared 2 days postinfection and immunoprecipitated with anti-EBNA1 antibody as in Fig. ​Fig.6.6. Immunoprecipitated proteins are shown. (B) Insect cell lysates coexpressing EBNA1 proteins and EBP2 (1/140 the amount used in panel A) were analyzed by Western blotting with EBNA1 antisera. The position of the band corresponding to the full-length EBNA1 protein in question is marked by the bracket.

Article Snippet: A baculovirus expressing the tagged EBP2 was generated by homologous recombination of pVLEBP2 with Baculogold baculovirus DNA (Pharmingen, San Diego, Calif.) as previously described ( 4 ).

Techniques: Expressing, Immunoprecipitation, Western Blot

Journal:

Article Title: EBP2, a Human Protein That Interacts with Sequences of the Epstein-Barr Virus Nuclear Antigen 1 Important for Plasmid Maintenance

doi:

Figure Lengend Snippet: Interaction of EBNA1 mutants with EBP2 in the two-hybrid system. (A) Activation of the HIS3 reporter was determined as in Fig. ​Fig.1.1. Tenfold serial dilutions of log-phase cultures of Y190 strains expressing EBNA1 or EBNA1 mutants as GAL4 DNA binding domain fusions and EBP2 or nothing (pACT2) fused to the GAL4 activation domain were plated on SC-Trp,Leu (left panel) or SC-Trp,Leu,His plus 50 mM AT (right panel). (B) Equal numbers of Y190 cells expressing the EBNA1 fusion proteins indicated were lysed and subjected to Western blot analysis with antibodies against the HA tag.

Article Snippet: A baculovirus expressing the tagged EBP2 was generated by homologous recombination of pVLEBP2 with Baculogold baculovirus DNA (Pharmingen, San Diego, Calif.) as previously described ( 4 ).

Techniques: Activation Assay, Expressing, Binding Assay, Western Blot

Journal:

Article Title: EBP2, a Human Protein That Interacts with Sequences of the Epstein-Barr Virus Nuclear Antigen 1 Important for Plasmid Maintenance

doi:

Figure Lengend Snippet: Summary of EBP2 interactions with EBNA1 fragments and mutants. A schematic representation of the EBNA1 polypeptides tested for EBP2 binding in co-immunoprecipitation (IP) and two-hybrid (2H) assays.

Article Snippet: A baculovirus expressing the tagged EBP2 was generated by homologous recombination of pVLEBP2 with Baculogold baculovirus DNA (Pharmingen, San Diego, Calif.) as previously described ( 4 ).

Techniques: Binding Assay, Immunoprecipitation