Journal: Cancer Science

Article Title: Carcinogenicity of dimethylarsinic acid in Ogg1 ‐deficient mice

doi: 10.1111/j.1349-7006.2007.00475.x

Figure Lengend Snippet: Gene Spring clustering analysis of the Affymetrix oligonucleotide microarray data (a) Ingenuity pathway analysis (b) and conformation of microarray results by real‐time quantitative polymerase chain reaction (Q‐PCR) (c–f). (c) Polymerase (DNA‐directed) alpha 1 (d) Mmp13 (e) nicotineamide adenine dinucleotide reduced form (NADH) dehydrogenase (ubiquinone) 1 alpha, and (f) CYP7b1 mRNA expression. *P < 0.05 and **P < 0.0001 versus Ogg1 +/+ control and dimethylarsinic acid (DMAV)‐treated groups.

Article Snippet: For the confirmation of Affimetrix microarray analysis results, real‐time quantitative‐polymerase chain reaction (Q‐PCR) was carried out using TaqMan probes for 490 C. Primer sequences were designed with Primer Express software (Applied Biosystems, USA): Mm00447106_m1 for polymerase (DNA‐directed), alpha 1 ( NM_008892 ); Mm00484157_m1 for cytochrome P450, 7b1 ( NM_007825 ); Mm00772875_m1 for nicotineamide adenine dinucleotide reduced form (NADH) dehydrogenase (ubiquinone) 1 alpha subcomplex, 3 ( NM_025348 ); Mm00439491_m1 for matrix metalloproteinase 13 ( NM_008607 ); Mm00834384_g1 for heat shock protein 1 ( NM_013560 ); and Mm00607939 for beta‐actin, cytoplasmic ( NM_007393 ).

Techniques: Microarray, Real-time Polymerase Chain Reaction, Expressing, Control

Journal: Inflammation Research

Article Title: Dexamethasone regulates gene expression in chondrocytes through MKP-1 and downregulates cholesterol hydroxylases CH25H and CYP7B1

doi: 10.1007/s00011-025-02121-5

Figure Lengend Snippet: CH25H and CYP7B1 expression was downregulated by dexamethasone in primary chondrocytes from wild-type (WT) mice, and the effect was attenuated in chondrocytes from MKP-1 deficient (knockout, KO) mice. Murine chondrocytes were stimulated with IL-1β (100 pg/mL), either alone or in combination with dexamethasone (1 μM), for 24 h. CH25H ( A ) and CYP7B1 ( B ) mRNA was measured by RT-PCR and normalized to GAPDH. For each genotype, IL-1β-treated samples were set as 100%, and the other values are presented in relation to those values. Data are shown as mean + SEM (n = 8). Statistical analysis was carried out with two-way ANOVA followed by Bonferroni post-test; ** p < 0.01, *** p < 0.0001

Article Snippet: Mouse CH25H (Mm00515486_s1), mouse CYP7B1 (Mm00484157_m1), human CH25H (Hs02379634_s1) and human CYP7B1 (Hs01046431_m1) were detected using TaqMan® Gene Expression assays obtained from Thermo Fisher Scientific.

Techniques: Expressing, Knock-Out, Reverse Transcription Polymerase Chain Reaction

Journal: Inflammation Research

Article Title: Dexamethasone regulates gene expression in chondrocytes through MKP-1 and downregulates cholesterol hydroxylases CH25H and CYP7B1

doi: 10.1007/s00011-025-02121-5

Figure Lengend Snippet: CH25H and CYP7B1 expressions were suppressed by dexamethasone in primary human OA chondrocytes. Chondrocytes were treated with interleukin-1β (IL-1β, 100 pg/mL), either alone or in combination with dexamethasone (1 μM), for 24 h. CH25H ( A ) and CYP7B1 ( B ) mRNA was measured by RT-PCR and normalized to GAPDH. Expression in the IL-1β-treated samples was set as 100%, and the other values are presented in relation to that value. Data are shown as mean + SEM (n = 5). Statistical analysis was carried out with repeated measures ANOVA followed by Bonferroni post-test; **** p < 0.0001

Article Snippet: Mouse CH25H (Mm00515486_s1), mouse CYP7B1 (Mm00484157_m1), human CH25H (Hs02379634_s1) and human CYP7B1 (Hs01046431_m1) were detected using TaqMan® Gene Expression assays obtained from Thermo Fisher Scientific.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Inflammation Research

Article Title: Dexamethasone regulates gene expression in chondrocytes through MKP-1 and downregulates cholesterol hydroxylases CH25H and CYP7B1

doi: 10.1007/s00011-025-02121-5

Figure Lengend Snippet: CH25H and CYP7B1 expressions were differently regulated by MAP kinase inhibitors. The primary human OA chondrocytes were treated with IL-1β (100 pg/mL) alone or in combination with the p38 MAP kinase inhibitor BIRB796 (100 nM) or with the JNK MAP kinase inhibitor SP600125 (10 μM) for 24 h. CH25H and CYP7B1 mRNA was measured by RT-PCR and normalized to GAPDH. CH25H ( A ) and CYP7B1 ( B ) expression levels in IL-1β -treated samples were set as 100%, and the other values are given in relation to those values. Data are shown as mean + SEM, n = 6. Statistical analysis was carried out with repeated measures ANOVA followed by Bonferroni post-test; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Mouse CH25H (Mm00515486_s1), mouse CYP7B1 (Mm00484157_m1), human CH25H (Hs02379634_s1) and human CYP7B1 (Hs01046431_m1) were detected using TaqMan® Gene Expression assays obtained from Thermo Fisher Scientific.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-1 Induces the Release of Lubricating Phospholipids from Human Osteoarthritic Fibroblast-Like Synoviocytes

doi: 10.3390/ijms23052409

Figure Lengend Snippet: IL-1ß induces the expression of cholesterol hydroxylase CH25H and CYP7B1 in FLS. CH25H ( A ) and CYP7B1 ( C ) were measured by Western blot or enzyme-linked immunosorbent assay ( B ). The mRNA expression of CH25H and CYP7B1 ( D ) was determined by reverse transcription-polymerase chain reaction (RT-PCR) using the 2 −ΔΔCt method. Representative western blots show the protein expression of cultured FLS as the corresponding control (−) and of FLS treated (+) with IL-1ß or T0901317. All western blots were quantified using ImageJ ( A , C ). Treated FLS and corresponding controls derived from the same experiment, and patient and gels/blots were processed in parallel. FLS were treated with 1 ng/mL IL-1ß or 0.1 µM T0901317 for 24, 48, and 72 h. Data for biological replicates represent fold-change versus corresponding controls (= 1, indicated by broken horizontal lines) and are shown as dot plots, with lines inside showing the mean ± SD ( n = 8). The y-axis represents the log of variable fold changes. Black circles show data from IL-1ß-treated FLS relative to untreated controls, whereas blue squares represent data from T0901317 relative to vehicle controls. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Equal amounts of proteins were subjected to SDS-PAGE on an 8% Tris-Glycine gel for ABCA1 and 10% gels for the remaining proteins and blotted onto PVDF membranes in a Trans-Blot ® SD semi-dry electrophoretic transfer cell (Bio-Rad Laboratories, Munich, Germany) at 1.0 A/cm 2 for 1.5 h. Membranes were blocked with buffered 5% skimmed milk for 1 h at room temperature and then incubated with primary antibodies against CH25H (#NBP2-83971), sterol regulatory element-binding protein 1c (SREBP-1c, gene SREBF1, clone 2A4, RRID: AB_10001575) (both from Novus Biologicals, Wiesbaden, Germany), CYP7B1 (clone OTI1G7, #TA807549, OriGene, Herford, Germany), ABCG1 (clone ARC0336, RRID:AB_2849090, Thermo Fisher), HMGCR (clone CL0260, RRID:AB_2786973, Invitrogen, Karlsruhe, Germany), liver X receptor alpha (LXRα; gene NR1H3, clone PPZ0412, RRID:AB_2154888), ABCA1 (clone 1276B, #MAB72071), APOE (#AF4144), GAPDH (#2275-PC-100, RRID: AB_2107456), and ß-actin (clone 937215, #MAB8929) (the last 5 from R&D Systems, Wiesbaden, Germany).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: Interleukin-1 Induces the Release of Lubricating Phospholipids from Human Osteoarthritic Fibroblast-Like Synoviocytes

doi: 10.3390/ijms23052409

Figure Lengend Snippet: Schematic of proposed mechanism of IL-1ß on phospholipid (PL) release from FLS during OA. IL-1ß induces the upregulation of cholesterol 25-hydroxylase (CH25H) and monooxygenase 25-hydroxycholesterol 7-alpha-hydroxylase, also called cytochrome P450 7B1 (CYP7B1), causing accumulation of oxysterols as endogenous ligands of nuclear liver X receptor (LXR). On activation of LXR by oxysterols or synthetic LXR agonists, such as T0901317, the protein level of active ATP-binding cassette transporter A1 (ABCA1) increases. The increased efflux of PLs to extracellular apolipoprotein (APO) A1 is mediated by ABCA1, which results in the formation of nascent HDL-c particles (nHDL). Oxysterols and T0901317 upregulate sterol regulatory element-binding protein (SREBP) 1c, suggesting that the biosynthesis of fatty acids that are needed for PLs is stimulated. Further, T0901317 enhances the expression of SREBP2, which binds as an important transcription factor to the promoter region of 3-hydroxy-3-methylglutaryl-coA reductase (HMGCR), the rate-limiting enzyme in cholesterol biosynthesis, to upregulate it. Dotted line: IL-1; solid line: T0901317; arrow: stimulation; dash: inhibition.

Article Snippet: Equal amounts of proteins were subjected to SDS-PAGE on an 8% Tris-Glycine gel for ABCA1 and 10% gels for the remaining proteins and blotted onto PVDF membranes in a Trans-Blot ® SD semi-dry electrophoretic transfer cell (Bio-Rad Laboratories, Munich, Germany) at 1.0 A/cm 2 for 1.5 h. Membranes were blocked with buffered 5% skimmed milk for 1 h at room temperature and then incubated with primary antibodies against CH25H (#NBP2-83971), sterol regulatory element-binding protein 1c (SREBP-1c, gene SREBF1, clone 2A4, RRID: AB_10001575) (both from Novus Biologicals, Wiesbaden, Germany), CYP7B1 (clone OTI1G7, #TA807549, OriGene, Herford, Germany), ABCG1 (clone ARC0336, RRID:AB_2849090, Thermo Fisher), HMGCR (clone CL0260, RRID:AB_2786973, Invitrogen, Karlsruhe, Germany), liver X receptor alpha (LXRα; gene NR1H3, clone PPZ0412, RRID:AB_2154888), ABCA1 (clone 1276B, #MAB72071), APOE (#AF4144), GAPDH (#2275-PC-100, RRID: AB_2107456), and ß-actin (clone 937215, #MAB8929) (the last 5 from R&D Systems, Wiesbaden, Germany).

Techniques: Activation Assay, Binding Assay, Expressing, Inhibition

Journal: PLoS ONE

Article Title: Fish Oil and the Pan-PPAR Agonist Tetradecylthioacetic Acid Affect the Amino Acid and Carnitine Metabolism in Rats

doi: 10.1371/journal.pone.0066926

Figure Lengend Snippet: Gene expression of selected genes in liver of rats after 50 weeks of diet administration 1 .

Article Snippet: Real-time PCR was performed with Sarstedt 384 well multiply-PCR Plates (Sarstedt Inc., Newton, NC, USA) on the following genes, using probes and primers from Applied Biosystems (Foster City, CA, USA): aminoadipate aminotransferase ( Aadat , Rn00567882_m1); aldehyde dehydrogenase 9 family, member 1 ( Aldh9a1 , Rn01491039_m1); aldehyde oxidase 3 ( Aox3 , Rn01441420_m1); arginase ( Arg1 , Rn00691090_m1); argininosuccinate lyase ( Asl , Rn01480437_g1); argininosuccinate synthetase ( Ass , Rn00565808_g1); gamma-butyrobetaine hydrolase 1 ( Bbox1 , Rn00575255_m1); cystathione-beta-synthase ( Cbs , Rn00560948_m1); ClpX caseinolytic peptidase×homolog (E.coli) ( Clpx , Rn01418137_m1); cytochrome P450, family 7, subfamily B, polypeptide 1 ( Cyp7b1 , Rn01461859_m1); glutamate dehydrogenase 1 ( Glud1 , Rn00561306_m1); glutamate-ammonia ligase ( Glul , Rn01483108_m1); glycerate kinase ( Glyctk , Rn01446603_g1); histidine decarboxylase ( Hdc , Rn00566665_m1); serine hydroxymethyltransferase ( Shmt2 , Rn01768052_g1); trimethyllysine hydroxylase, epsilon ( Tmlhe , Rn00591314_m1).

Techniques: Gene Expression

Journal: Orphanet Journal of Rare Diseases

Article Title: Chenodeoxycholic acid rescues axonal degeneration in induced pluripotent stem cell-derived neurons from spastic paraplegia type 5 and cerebrotendinous xanthomatosis patients

doi: 10.1186/s13023-023-02666-w

Figure Lengend Snippet: Establishment and characterization of SPG5 iPSCs. a Phase-contrast images of SPG5 dermal fibroblasts and iPSCs. b PCR gel images showing the expression of FGF5 , NANOG , SOX2 , and OCT4 in SPG5 fibroblasts (FB) and iPSCs. GAPDH is a housekeeping gene. c and d The mRNA expression of OCT4 and FGF5 in fibroblasts and iPSCs by qRT-PCR. e Immunostaining showing the protein expression of pluripotency markers NANOG, SSEA4, and TRA-1-60 (red) in iPSCs. Blue: Hoechst. f No expression of pluripotency markers was detected in SPG5 fibroblasts. Blue: Hoechst. g Genomic DNA sequencing confirmed the compound heterozygous mutations c.334C > T, p.Arg112* and c.1456C > T, p.Arg486Cys in the CYP7B1 gene. Data are represented as means ± SD. * p < 0.05 compared to fibroblast cells by two-sided Student’s t -test. Scale bars, 100 µm ( a ) and 50 µm ( e and f )

Article Snippet: To knockout (KO) CYP7B1 in hESCs, CYP7B1 CRISPR-cas9 KO plasmids containing RNA sequences that specifically target the CYP7B1 gene were obtained from Santa Cruz Biotechnology (Cat. #: sc-405345).

Techniques: Expressing, Quantitative RT-PCR, Immunostaining, DNA Sequencing

Journal: Orphanet Journal of Rare Diseases

Article Title: Chenodeoxycholic acid rescues axonal degeneration in induced pluripotent stem cell-derived neurons from spastic paraplegia type 5 and cerebrotendinous xanthomatosis patients

doi: 10.1186/s13023-023-02666-w

Figure Lengend Snippet: CDCA treatment rescued axonal degeneration induced by loss of CYP7B1. a Representative phase-contrast image showing CYP7B1 KO stem cell clones generated via CRISPR-cas9-mediated gene editing of hESCs. These ESCs were then differentiated into neurons. Scale bar, 50 µm. b qPCR showing mRNA expression of CYP7B1 in H9, CYP7B1 KO #a, and CYP7B1 KO #b cortical PNs. c Double immunostaining of TAU and CTIP2 in the indicated cortical PNs. Red: CTIP2; green: TAU; blue: Hoechst. Scale bar, 50 µm. d Axonal outgrowth quantifications of the indicated cortical PNs. e Quantifications of NFL expression in the indicated PNs. f Representative images showing immunostaining of TAU in H9, CYP7B1 KO #a, and CYP7B1 KO #b cortical PNs after vehicle or CDCA treatment. Axonal swellings are magnified and indicated with arrowheads. Scale bar, 20 µm. g A representative image showing double immunostaining of TAU and pNFH in CYP7B1 KO cortical PN axon swellings (arrowheads). Scale bar, 5 µm. h Quantification of TAU + axonal swellings in H9, CYP7B1 KO #a, and CYP7B1 KO #b cortical PNs after vehicle or CDCA treatment. Data are represented as means ± SEM. * p < 0.05 compared to H9 neurons treated with vehicle by Dunnett’s test after ANOVA ( b , d , e, and h ). # p < 0.05 compared to CYP7B1 KO vehicle-treated group by two-sided Student t -test ( h )

Article Snippet: To knockout (KO) CYP7B1 in hESCs, CYP7B1 CRISPR-cas9 KO plasmids containing RNA sequences that specifically target the CYP7B1 gene were obtained from Santa Cruz Biotechnology (Cat. #: sc-405345).

Techniques: Clone Assay, Generated, CRISPR, Expressing, Double Immunostaining, Immunostaining

Journal: Orphanet Journal of Rare Diseases

Article Title: Chenodeoxycholic acid rescues axonal degeneration in induced pluripotent stem cell-derived neurons from spastic paraplegia type 5 and cerebrotendinous xanthomatosis patients

doi: 10.1186/s13023-023-02666-w

Figure Lengend Snippet: Establishment and characterization of SPG5 iPSCs. a Phase-contrast images of SPG5 dermal fibroblasts and iPSCs. b PCR gel images showing the expression of FGF5 , NANOG , SOX2 , and OCT4 in SPG5 fibroblasts (FB) and iPSCs. GAPDH is a housekeeping gene. c and d The mRNA expression of OCT4 and FGF5 in fibroblasts and iPSCs by qRT-PCR. e Immunostaining showing the protein expression of pluripotency markers NANOG, SSEA4, and TRA-1-60 (red) in iPSCs. Blue: Hoechst. f No expression of pluripotency markers was detected in SPG5 fibroblasts. Blue: Hoechst. g Genomic DNA sequencing confirmed the compound heterozygous mutations c.334C > T, p.Arg112* and c.1456C > T, p.Arg486Cys in the CYP7B1 gene. Data are represented as means ± SD. * p < 0.05 compared to fibroblast cells by two-sided Student’s t -test. Scale bars, 100 µm ( a ) and 50 µm ( e and f )

Article Snippet: To knockout (KO) CYP7B1 in hESCs, CYP7B1 CRISPR-cas9 KO plasmids containing RNA sequences that specifically target the CYP7B1 gene were obtained from Santa Cruz Biotechnology (Cat. #: sc-405345).

Techniques: Expressing, Quantitative RT-PCR, Immunostaining, DNA Sequencing

Journal: Orphanet Journal of Rare Diseases

Article Title: Chenodeoxycholic acid rescues axonal degeneration in induced pluripotent stem cell-derived neurons from spastic paraplegia type 5 and cerebrotendinous xanthomatosis patients

doi: 10.1186/s13023-023-02666-w

Figure Lengend Snippet: CDCA treatment rescued axonal degeneration induced by loss of CYP7B1. a Representative phase-contrast image showing CYP7B1 KO stem cell clones generated via CRISPR-cas9-mediated gene editing of hESCs. These ESCs were then differentiated into neurons. Scale bar, 50 µm. b qPCR showing mRNA expression of CYP7B1 in H9, CYP7B1 KO #a, and CYP7B1 KO #b cortical PNs. c Double immunostaining of TAU and CTIP2 in the indicated cortical PNs. Red: CTIP2; green: TAU; blue: Hoechst. Scale bar, 50 µm. d Axonal outgrowth quantifications of the indicated cortical PNs. e Quantifications of NFL expression in the indicated PNs. f Representative images showing immunostaining of TAU in H9, CYP7B1 KO #a, and CYP7B1 KO #b cortical PNs after vehicle or CDCA treatment. Axonal swellings are magnified and indicated with arrowheads. Scale bar, 20 µm. g A representative image showing double immunostaining of TAU and pNFH in CYP7B1 KO cortical PN axon swellings (arrowheads). Scale bar, 5 µm. h Quantification of TAU + axonal swellings in H9, CYP7B1 KO #a, and CYP7B1 KO #b cortical PNs after vehicle or CDCA treatment. Data are represented as means ± SEM. * p < 0.05 compared to H9 neurons treated with vehicle by Dunnett’s test after ANOVA ( b , d , e, and h ). # p < 0.05 compared to CYP7B1 KO vehicle-treated group by two-sided Student t -test ( h )

Article Snippet: To knockout (KO) CYP7B1 in hESCs, CYP7B1 CRISPR-cas9 KO plasmids containing RNA sequences that specifically target the CYP7B1 gene were obtained from Santa Cruz Biotechnology (Cat. #: sc-405345).

Techniques: Clone Assay, Generated, CRISPR, Expressing, Double Immunostaining, Immunostaining