Journal: The Journal of Cell Biology

Article Title: Spindle assembly checkpoint robustness requires Tpr-mediated regulation of Mad1/Mad2 proteostasis

doi: 10.1083/jcb.201309076

Figure Lengend Snippet: Tpr is required for SAC proteostasis throughout the cell cycle. (A and B) Normalized expression of tpr , mad1 , mad2 , mps1 , cdc20 , and p31 in control and Tpr-depleted asynchronous or mitotic cells. Error bars represent standard deviations from three independent experiments. (C) WB analysis of asynchronous, G2, and mitotic enriched HeLa cell extracts from control (+) and Tpr-depleted (−) cells with (+) or without (−) MG132 with the indicated antibodies. The percentage of protein levels relative to controls is indicated. (D) c-Mad2 IP from asynchronous and mitotic HeLa cells with (+) or without (−) Tpr, in the presence (+) or absence (−) of MG132. (E and F) Cells with or without Tpr were treated with cycloheximide (CHX) for various time points as indicated. Total protein extracts of asynchronous HeLa cells were analyzed by WB to detect Tpr, Mps1, Mad1, and t-Mad2 in control and Tpr-depleted cells. α-Tubulin was used as a loading control. *, P < 0.05; **, P < 0.01; ***, P < 0.001. The error bars indicate standard deviations from two independent experiments. (G) Immunodetection of Mad1 (red), SENP1, and SENP2 (green) in Tpr-depleted cells (−). A nondepleted cell (+) was used as internal control. DNA was counterstained with DAPI (blue). (H) WB analysis of asynchronous cell extracts in control and after SENP1 or SENP2 RNAi. The percentage of protein levels relative to controls is indicated. α-Tubulin was used as a loading control. Bar, 10 µm.

Article Snippet: Mouse anti-Mad1 (generated against full-length Mad1, 1:500; provided by A. Santamaria and E. Nigg, Biozentrum, Basel, Switzerland), rabbit anti-Mad1 (generated against full-length Mad1, 1:1,000; provided by P. Meraldi, University of Geneva, Switzerland), mouse anti–c-Mad2 (generated against full-length Mad2, 1:500; provided by A. Santamaria), rabbit anti–t-Mad2 (1:300; Bethyl Laboratories, Inc.), sheep anti–o-Mad2 (generated against full-length Mad2, 1:200; provided by S. Taylor, University of Manchester, UK), rabbit anti-Cdc20 (1:100; Santa Cruz Biotechnology, Inc.), mouse anti-Mps1 (1:100; Merck Millipore), rabbit anti-Tpr (1:500; Novus Biologicals), sheep anti-BubR1 (generated against aa 2–422, 1:300; provided by S. Taylor), rabbit anti-SENP1 or -SENP2 (generated against aa 273–449 and aa 1–92, respectively; 1:1,000; provided by M. Dasso, National Institutes of Health, Bethesda, MD), and human anticentromere antibodies (ACAs; 1:5,000, provided by B. Earnshaw; or 1:2,000, Fitzgerald Industries International) were used as primary antibodies, and Alexa Fluor 488, 568, and 647 (Invitrogen) were used as secondary antibodies (1:1,000).

Techniques: Expressing, Control, Immunodetection

Journal: Clinical and Developmental Immunology

Article Title: PADI4 Haplotypes in Association with RA Mexican Patients, a New Prospect for Antigen Modulation

doi: 10.1155/2013/383681

Figure Lengend Snippet: Genotyping strategies for PADI4 polymorphism variants detection.

Article Snippet: The PADI4_89, PADI4_90, and PADI4_92 genotypes were identified after restriction enzyme digestion with HaeIII , MscI, and MspI, respectively (New England Biolabs, MS, USA), shown in .

Techniques: Sequencing

Journal: Clinical and Developmental Immunology

Article Title: PADI4 Haplotypes in Association with RA Mexican Patients, a New Prospect for Antigen Modulation

doi: 10.1155/2013/383681

Figure Lengend Snippet: PADI4 SNPs enzyme digestion. The figure shows digestion of three SNPs in the PADI4 gene. (a) Shows digestion of PADI4_89, with HaeIII enzyme; lane 1 represents the A/A genotype, lane 2 A/G and 3 G/G. (b) Demonstrates PADI4_90 amplification (221 bp) in lane 1 and digested products with MscI enzyme in lanes 2 (C/C genotype), 3 (C/T genotype), and 4 (T/T genotype). (c) Shows amplification product of PADI4_92 in lane 1 (363 bp) and restriction products obtained with the enzyme MspI; lane 2 corresponds to the G/G genotype, lane 3 G/C, and lane 4 C/C. Visualized in 8% (29 : 1) polyacrylamide gel with silver staining. M: molecular weight marker (50 bp).

Article Snippet: The PADI4_89, PADI4_90, and PADI4_92 genotypes were identified after restriction enzyme digestion with HaeIII , MscI, and MspI, respectively (New England Biolabs, MS, USA), shown in .

Techniques: Amplification, Silver Staining, Molecular Weight, Marker

Journal: Clinical and Developmental Immunology

Article Title: PADI4 Haplotypes in Association with RA Mexican Patients, a New Prospect for Antigen Modulation

doi: 10.1155/2013/383681

Figure Lengend Snippet: Genotypic and allelic frequencies of PADI4_89, PADI4_90, and PADI4_92 SNPs of PADI4 gene in controls (HC n = 98) and rheumatoid arthritis (RA n = 86) patients.

Article Snippet: The PADI4_89, PADI4_90, and PADI4_92 genotypes were identified after restriction enzyme digestion with HaeIII , MscI, and MspI, respectively (New England Biolabs, MS, USA), shown in .

Techniques:

Journal: Clinical and Developmental Immunology

Article Title: PADI4 Haplotypes in Association with RA Mexican Patients, a New Prospect for Antigen Modulation

doi: 10.1155/2013/383681

Figure Lengend Snippet: Haplotype sequence and frequency of PADI4 gene SNPs (PADI4_89, PADI4_90, and PADI4_92) in healthy controls and RA patients.

Article Snippet: The PADI4_89, PADI4_90, and PADI4_92 genotypes were identified after restriction enzyme digestion with HaeIII , MscI, and MspI, respectively (New England Biolabs, MS, USA), shown in .

Techniques: Sequencing

Journal: Clinical and Developmental Immunology

Article Title: PADI4 Haplotypes in Association with RA Mexican Patients, a New Prospect for Antigen Modulation

doi: 10.1155/2013/383681

Figure Lengend Snippet: Anti-cyclic citrullinated peptide antibodies (ACPA) titers in the presence of genetic variants in PADI4_89, PADI4_90, and PADI4_92 polymorphisms of PADI4 gene.

Article Snippet: The PADI4_89, PADI4_90, and PADI4_92 genotypes were identified after restriction enzyme digestion with HaeIII , MscI, and MspI, respectively (New England Biolabs, MS, USA), shown in .

Techniques:

Journal: Clinical and Developmental Immunology

Article Title: PADI4 Haplotypes in Association with RA Mexican Patients, a New Prospect for Antigen Modulation

doi: 10.1155/2013/383681

Figure Lengend Snippet: Association studies for PADI4 SNPs and rheumatoid arthritis.

Article Snippet: The PADI4_89, PADI4_90, and PADI4_92 genotypes were identified after restriction enzyme digestion with HaeIII , MscI, and MspI, respectively (New England Biolabs, MS, USA), shown in .

Techniques: Expressing