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: PLOS One

Article Title: Myeloid Fmr1 deficiency in mice results in reduced serum cholesterol and altered bile pathway gene expression

doi: 10.1371/journal.pone.0340222

Figure Lengend Snippet: Tgr5 mRNA (A) expression in liver of myeloid-specific Fmr1 deficient (Fmr1 fl -LysM cre ) compared to control (Fmr1 fl ), N = 5 each. Representative Western blot (B) of Tgr5 from liver extract of Fmr1 fl -LysM cre and Fmr1 fl mice with quantification below, N = 4-5 each. IL-1β (C) and IL-6 (D) mRNA expression in livers (N = 4-5 each). (A-D) *P < 0.05 ANOVA with multiple comparison test. BMDM Tgr5 mRNA expression in male; N = 4-5 each (E) and Western blot with quantification below (F) in both female (N = 2 each) and male (N = 2-3) Fmr1 fl compared to Fmr1 fl -LysM cre ; **P < 0.01 T test. Gating scheme (G) for Tgr5 staining and FACS analysis of peritoneal macrophage double positive for F4/80 and CD11b. Percentage of Tgr5 + peritoneal macrophage (H) of Fmr1 fl (N = 8; 4 each sex) compared to Fmr1 fl -LysM cre (N = 6; 3 each sex) mice. *P < 0.05 T test.

Article Snippet: Primary antibodies used were: LDLR (Invitrogen), Cyp7b1 (Proteintech), Cyp27a1 (Abcam) and Tgr5 (Proteintech).

Techniques: Expressing, Control, Western Blot, Comparison, Staining

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: Molecular Therapy. Methods & Clinical Development

Article Title: AAV8-based gene replacement therapy for hereditary spastic paraplegia type 5

doi: 10.1016/j.omtm.2025.101531

Figure Lengend Snippet: In vitro validation of generated constructs (A) Overview of in vitro experiments. (B) Gene expression of CYP7B1 in HepG2 cells after transfection with plasmids for GFP (control), hCYP7B1, and CYP7B1-FLAG assessed by qPCR. CYP7B1 expression after transfection with the respective plasmids is upregulated 10,000-fold compared to untreated and GFP controls. (C) Western blot analysis confirms strong expression of CYP7B1 (VCL, vinculin, loading control). (D) Immunofluorescence of HepG2 cells fixed 24 h after transfection with the CYP7B1-FLAG construct, stained against the FLAG tag, the ER marker CLIMP63, and DNA (scale bars, 20 μm).

Article Snippet: Immunohistochemical staining for CYP7B1 (Proteintech, 24889-1-AP) was performed on 4 μm deparaffinized sections.

Techniques: In Vitro, Biomarker Discovery, Generated, Construct, Gene Expression, Transfection, Control, Expressing, Western Blot, Immunofluorescence, Staining, FLAG-tag, Marker

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: AAV8-based gene replacement therapy for hereditary spastic paraplegia type 5

doi: 10.1016/j.omtm.2025.101531

Figure Lengend Snippet: Liver transduction efficacy and liver oxysterol levels (A) Immunohistochemistry of liver sections against CYP7B1 from wild-type (WT) animals, Cyp7B1 −/− animals injected with different doses of AAV8-TTR-hCYP7B1, AAV8-TTR-eGFP (vehicle), and untreated KO animals (KO). (B) mRNA levels of human CYP7B1 and endogenous murine Cyp7b1 for the six experimental groups normalized to Eif2α and Actβ. Strong expression of human CYP7B1 is observed with all doses of AAV-TTR-hCYP7B1. (C) Western blot of liver homogenates (VCL, vinculin, loading control). CYP7B1 expression in all groups receiving AAV-TTR-CYP7B1 injections exceeded WT expression and highest doses led to oversaturation of the membrane, while untreated and vehicle-injected KO animals show no signal for CYP7B1. (D) Liver oxysterol levels at the endpoint of the study (D49). Both 25-HC and 27-HC were significantly reduced compared to the GFP vehicle-treated group. Levels of the cerebral metabolite 24-HC decreased slightly but remained within the range of WT animals. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; and ∗∗∗∗ p < 0.0001.

Article Snippet: Immunohistochemical staining for CYP7B1 (Proteintech, 24889-1-AP) was performed on 4 μm deparaffinized sections.

Techniques: Transduction, Immunohistochemistry, Injection, Expressing, Western Blot, Control, Membrane