Journal: Drug Metabolism and Disposition

Article Title: Impact of the transcription factor nuclear factor 1 B T>C polymorphism on clozapine metabolism in vivo and expression of intestinal transporters in vitro

doi: 10.1016/j.dmd.2025.100100

Figure Lengend Snippet: Effect of NFIB rs28379954 T>C diplotype on metabolic ratios in smoking patients adjusting for age, sex, and sampling time. The data are analyzed using linear mixed model. The figure displays the fold change (95% CIs) in NFIB CT carriers (heterozygous for the target allele) versus NFIB TT carriers (homozygous for the target allele). Significant effect was observed for the metabolic ratio of N -desmethylclozapine cysteine: DCL-CYS/DCL (fold change: 1.89; CI: 1.04–3.44; P = .0384). contains the effect estimates, fold changes, and 95% CIs of the rest of the metabolites. CI, confidence interval; DCL, N -desmethylclozapine; DCL-CYS/DCL, DCL-cysteinyl; NFIB, nuclear factor 1 B.

Article Snippet: A predesigned TaqMan-based real-time polymerase chain reaction assay NFIB rs28379954 T>C (Thermo Fisher Scientific) was used for the determination of NFIB genotype.

Techniques: Sampling

Journal: Drug Metabolism and Disposition

Article Title: Impact of the transcription factor nuclear factor 1 B T>C polymorphism on clozapine metabolism in vivo and expression of intestinal transporters in vitro

doi: 10.1016/j.dmd.2025.100100

Figure Lengend Snippet: Effect of NFIB rs28379954 T>C diplotype on metabolic ratios in nonsmoking patients adjusting for age, sex, and sampling time. The data are analyzed using linear mixed model. The figure displays the fold change (95% CIs) in NFIB CT carriers (heterozygous for the target allele) versus NFIB TT carriers (homozygous for the target allele). No significant effect was observed for the metabolic ratios of the 30 metabolites. contains the effect estimates, fold changes, and 95% CIs of all the metabolites. CI, confidence interval; NFIB, nuclear factor 1 B.

Article Snippet: A predesigned TaqMan-based real-time polymerase chain reaction assay NFIB rs28379954 T>C (Thermo Fisher Scientific) was used for the determination of NFIB genotype.

Techniques: Sampling

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Molecular Basis for Dysregulated Activation of NKX 2‐5 in the Vascular Remodeling of Systemic Sclerosis

doi: 10.1002/art.40419

Figure Lengend Snippet: Binding of transcription‐enhancer factor domain 1 ( TEAD 1) at the rs3095870 site and increased NKX 2-5 transcriptional activity. Binding assays were performed using nuclear extracts ( NE ) from transforming growth factor β–treated immortalized human pulmonary artery smooth muscle cells ( PASMC s) and biotinylated DNA probes containing the rs3095870 C/T alleles ( A–C ). A, Electrophoretic mobility shift assay with rs3095870 C allele biotinylated probe (lane 2), and supershifts with TEAD 1 and Yes‐associated protein 1 ( YAP 1) antibodies (Ab) (lanes 3 and 4), as well as without nuclear extract (lane 1). B, Pull‐down assay using streptavidin beads and biotinylated DNA probes specific to rs3095870 C/T alleles. Complexes were analyzed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and Western blotting and immunodecorated with TEAD 1 antibody. Nonbiotinylated and scrambled (scr) biotinylated probes were used as controls. Input was 5% of total nuclear extract. C, Chromatin immunoprecipitation assays using antibodies specific for TEAD 1, phosphorylated Smad3 (Ph‐Smad3), and RNA polymerase II ( RNA Pol II ). Specific polymerase chain reaction primers were designed and used to detect enrichment. Immunoprecipitation with IgG was used as a control. No template control ( NTC ) was also used. Input was 10% of the initial chromatin used per immunoprecipitation assay. A schematic representation of the NKX 2-5 gene, showing intron 1, exons 1 and 2, and untranslated regions (3′‐ UTR and 5′‐ UTR ), as well as the locations of the minimal promoter and rs3095870 C>T site, is shown at the top. D, Luciferase reporter gene assays of primary human PASMC s. TEAD 1 expression vector was cotransfected with DNA constructs containing either the minimal promoter ( pGL ‐minprom) or the upstream promoter with the rs3095870‐C/T alleles ( pGL ‐prom‐C/T). The pGL 4.10 vector, which contains the firefly luciferase gene, was used as the control. Values are the mean ± SEM of 3 independent experiments. **** = P ≤ 0.0001 by Student's t ‐test.

Article Snippet: For rs3095870, a high‐resolution melting analysis was performed using a Type‐it high‐resolution melting polymerase chain reaction (PCR) kit (Qiagen).

Techniques: Binding Assay, Activity Assay, Electrophoretic Mobility Shift Assay, Pull Down Assay, Polyacrylamide Gel Electrophoresis, Western Blot, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Immunoprecipitation, Luciferase, Expressing, Plasmid Preparation, Construct