Journal: Journal of Bone and Mineral Research

Article Title: Effects of 25-Hydroxyvitamin D 3 on Proliferation and Osteoblast Differentiation of Human Marrow Stromal Cells Require CYP27B1/1α-Hydroxylase

doi: 10.1002/jbmr.298

Figure Lengend Snippet: Primer Sets Used for RT-PCR

Article Snippet: Transient transfection of siRNA into hMSCs hi-1α was performed by electroporation with the Human MSC Nucleofector Kit (Lonza/Amaxa Biosystems, Walkersville, MD, USA) with either CYP27B1 siRNA, nonsilencing control siRNA (a nonhomologous, scrambled sequence equivalent; Santa Cruz Biotechnology, Inc.), or PBS according to the manufacturer's instructions and as described previously. ( ) In brief, hMSCs hi-1α were harvested by trypsinization and resuspended at 10 6 cells in 100 μL of Nucleofector Solution (Lonza/Amaxa Biosystems) with 10 or 100 pmol of CYP27B1 siRNA.

Techniques: Sequencing

Journal: Nature Communications

Article Title: CYP6AE gene cluster knockout in Helicoverpa armigera reveals role in detoxification of phytochemicals and insecticides

doi: 10.1038/s41467-018-07226-6

Figure Lengend Snippet: Metabolism of xanthotoxin, 2-tridecanone, and indoxacarb by recombinant P450s. CYP6AE subfamily enzymes were from H. armigera and CYP6B1 from Papilio polyxenes . Error bars represent mean values ± SEM ( n = 4). Asterisk indicates that no significant metabolism was detected (limits of detection were 0.081, 0.033, and 0.016 pmol/min/pmol P450 for xanthotoxin, 2-tridecanone, and indoxacarb, respectively)

Article Snippet: In vitro metabolism was performed with 20 pmol recombinant P450, NADPH regeneration system (Promega, Madison, WI) and 2 μl substrate (20 μM xanthotoxin dissolved in DMSO, 20 μM 2-tridecanone or 10 μM indoxacarb dissolved in acetonitrile) in 200 μl 0.1 M potassium phosphate buffer (pH 7.4).

Techniques: Recombinant

Journal: Environment international

Article Title: Genome-modified Caenorhabditis elegans expressing the human cytochrome P450 (CYP1A1 and CYP1A2) pathway: An experimental model for environmental carcinogenesis and pharmacological research.

doi: 10.1016/j.envint.2024.109187

Figure Lengend Snippet: Fig. 2. BPDE-DNA adducts measurement by ELISA. (A) BPDE-DNA adducts were quantified in the two double-humanized C. elegans (CYP1A1_EPHX and CYP1A2_EPHX) and the wild-type (N2) worm exposed to 40 and 640 μM BaP for 48 h. Values represent mean ± SEM (n = 4 ± 1). Statistical analysis was performed via a two-way ANOVA, followed by a Turkey’s multiple comparison test, *p < 0.05, ***p < 0.001. (B) Determination of dG-N2-BPDE adduct formation by 32P- postlabelling for N2 (I), CYP1A1;EPHX (II) and CYP1A2;EPHX (III) exposed to 40 and 640 μM BaP for 48 h. The standards were prepared by incubating calf thymus DNA with 1 µM BaP which served as a positive control. Autoradiography was conducted for 24 h.

Article Snippet: Recombinant CYP1A1 protein (0.005 pmol) served as positive control (Corning® SupersomesTM Human CYP1A1 + Oxidoreductase, 0.5 nmol) and the CYP1A1 polyclonal antibody (1:2000, Proteintech, 13241-1-AP) was utilised as first antibody and the anti-mouse immunoglobulins (1:10000) as secondary antibody.

Techniques: Enzyme-linked Immunosorbent Assay, Comparison, Positive Control, Autoradiography

Journal: Environment international

Article Title: Genome-modified Caenorhabditis elegans expressing the human cytochrome P450 (CYP1A1 and CYP1A2) pathway: An experimental model for environmental carcinogenesis and pharmacological research.

doi: 10.1016/j.envint.2024.109187

Figure Lengend Snippet: Fig. 3. (A) Expression profile of CYP1A1;EPHX and CYP1A2;EPHX and the background strain 6070_6401 exposed to DMSO only, 40 or 640 μM BaP. The cluster analysis results of the fragments per kilobase of transcript per million mapped reads (FPKM) of three biological replicates for each condition utilised log2(FPKM + 1) values. The red colour indicates genes with high expression levels, and blue colour represents genes with low expression levels. The phylogenetic tree was constructed using the Euclidean distance algorithm. (B) Bar graph summarising the number of the significantly (p < 0.05) upregulated (red) and downregulated (blue) genes in worms exposed to 40 and 640 μM BaP compared to DMSO as control in each strain. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Recombinant CYP1A1 protein (0.005 pmol) served as positive control (Corning® SupersomesTM Human CYP1A1 + Oxidoreductase, 0.5 nmol) and the CYP1A1 polyclonal antibody (1:2000, Proteintech, 13241-1-AP) was utilised as first antibody and the anti-mouse immunoglobulins (1:10000) as secondary antibody.

Techniques: Expressing, Construct, Control

Journal: Environment international

Article Title: Genome-modified Caenorhabditis elegans expressing the human cytochrome P450 (CYP1A1 and CYP1A2) pathway: An experimental model for environmental carcinogenesis and pharmacological research.

doi: 10.1016/j.envint.2024.109187

Figure Lengend Snippet: Fig. 6. (A) Principal component analysis of the whole genome sequencing data of the single nucleotide variant (SNV) of CYP1A1;EPHX and CYP1A2;EPHX and the wild-type worm exposed to 640 μM BaP. All samples contained DMSO (0.5 %, v/v). (B) Bar plot representing the mean occurrence of each type of SNV in each strain exposed to 640 µM BaP. The values denote mean ± SEM (n = 3). ***p < 0.001, **p < 0.01, *p < 0.05.

Article Snippet: Recombinant CYP1A1 protein (0.005 pmol) served as positive control (Corning® SupersomesTM Human CYP1A1 + Oxidoreductase, 0.5 nmol) and the CYP1A1 polyclonal antibody (1:2000, Proteintech, 13241-1-AP) was utilised as first antibody and the anti-mouse immunoglobulins (1:10000) as secondary antibody.

Techniques: Sequencing, Variant Assay

Journal: Analytical and Bioanalytical Chemistry

Article Title: Digital microfluidic immobilized cytochrome P450 reactors with integrated inkjet-printed microheaters for droplet-based drug metabolism research

doi: 10.1007/s00216-018-1280-7

Figure Lengend Snippet: a Resorufin calibration curve and schematics of the fluorescence quantification by well-plate reader. b The fluorescence signal ( λ ex = 570 nm, λ em = 590 nm) of the reaction solutions (positive controls, n = 10, and negative controls, n = 4 CYP-IMERs) extracted from the monolith-based rCYP1A1-IMERs and analyzed on chip with the well-plate reader

Article Snippet: 0.88 ± 0.39 fmol ( n = 10 CYP-IMERs), assuming that the enzyme activity of the immobilized rCYP1A1 is similar to that of the soluble rCYP1A1 (23 pmol/min/pmol P450, Corning).

Techniques: Fluorescence