Journal: Experimental & Molecular Medicine

Article Title: Disruption of cholesterol homeostasis triggers periodontal inflammation and alveolar bone loss

doi: 10.1038/s12276-023-01122-w

Figure Lengend Snippet: Representative images of filipin staining of gingival tissue from human patients with PD ( a ) and in the maxilla region of ligature-induced PD mice ( b ). Scale bar, 25 μm. Total, free cholesterol, and cholesteryl ester ( n = 6) levels ( c ) and mRNA ( n = 3) ( d ) levels of cholesterol synthesis-related genes in human GFs treated with IL1β or TNFα. e Total cholesterol level in human GFs treated with the cholesterol synthesis inhibitors triparanol (TP) or lovastatin (LV) in the presence of IL1β (2 ng ml −1 ) or TNFα (50 ng ml −1 ) for 24 h ( n = 4). f mRNA levels of MMP1 , MMP3 , IL6 , IL8 , CCL5 , and PTGS2 in human GFs treated with cholesterol synthesis inhibitors (TP and LV) in the presence of IL1β (2 ng ml −1 ) or TNFα (50 ng ml −1 ) for 24 h ( n = 4). n indicates the number of biologically independent samples. Values are presented as the mean ± SEM based on one-way ANOVA with Tukey’s test. (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: For determination of cellular cholesterol levels, human GFs were treated with IL1β, TNFα, cholesterol synthesis inhibitors (triparanol (Cayman) and lovastatin (Enzo), or anti-CXCL16 blocking antibody (R&D Systems) in the presence of IL1β (2 ng ml −1 ), TNFα (50 ng ml −1 ) or cholesterol (200 μM).

Techniques: Staining

Journal: Experimental & Molecular Medicine

Article Title: Disruption of cholesterol homeostasis triggers periodontal inflammation and alveolar bone loss

doi: 10.1038/s12276-023-01122-w

Figure Lengend Snippet: a Representative images and measurement of Dil-OxLDL uptake into human GFs treated with or without IL1β (2 ng ml −1 ) or TNFα (50 ng ml −1 ) for 24 h ( n = 5). Scale bar, 25 μm. b Transcript levels of cholesterol uptake receptors in human GFs treated with IL1β (5 ng ml −1 ) or TNFα (100 ng ml −1 ) ( n = 3). Representative images of CXCL16 immunostaining of the gingiva from human patients with PD ( c ) and ligature-induced PD mice ( d ). Total cholesterol levels ( n = 4) ( e ) and measurement of Dil-OxLDL uptake into human GFs ( n = 5) ( f ) treated with an anti-CXCL16 blocking antibody in the presence of IL1β (2 ng ml −1 ) or TNFα (50 ng ml −1 ) for 24 h. n indicates the number of biologically independent samples. Values are presented as the mean ± SEM based on a two-tailed t -test ( a ) and one-way ANOVA with Tukey’s test ( b , e , f ). (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: For determination of cellular cholesterol levels, human GFs were treated with IL1β, TNFα, cholesterol synthesis inhibitors (triparanol (Cayman) and lovastatin (Enzo), or anti-CXCL16 blocking antibody (R&D Systems) in the presence of IL1β (2 ng ml −1 ), TNFα (50 ng ml −1 ) or cholesterol (200 μM).

Techniques: Immunostaining, Blocking Assay, Two Tailed Test

Journal: Experimental & Molecular Medicine

Article Title: Disruption of cholesterol homeostasis triggers periodontal inflammation and alveolar bone loss

doi: 10.1038/s12276-023-01122-w

Figure Lengend Snippet: a Heatmaps of genes involved in cholesterol metabolism and metabolic pathways of bile acid synthesis from the microarray analysis of human GFs treated with IL1β (left square) and TNFα (right circle) ( n = 3). Transcript levels of CH25H and CYP7B1 in human GFs treated with IL1β (5 ng ml −1 ) and TNFα (100 ng ml −1 ) for 24 h ( n = 4) ( b ) and the gingiva of healthy patients and patients with PD ( n = 6) ( c ). d Representative immunostaining images of CH25H and CYP7B1 in human and mouse gingiva. Scale bar, 25 μm. e , f mRNA levels in human GFs infected with empty (Ad-C) or CH25H -encoding adenovirus (Ad- CH25H ) ( n = 3) for 48 h ( n = 3). g Protein levels of CH25H, MMP1, IL6, IL8, and PTGS2 in cell lysates of human GFs infected with CH25H -overexpressing adenovirus (Ad- CH25H ; n = 3). h , i Representative images of CH25H immunostaining, μCT, and H&E staining images of mouse gingiva injected with an empty virus (Ad-C) or Ad- Ch25h (1 × 10 9 PFU per 5 μl). Magnification, ×100. Scale bar, 100 μm (left). Scale bar, 25 μm (right) ( h ). Magnification, ×200. Scale bar, 100 μm ( i ). j BMD analysis and CEJ-ABC distance measurement of the mouse maxillae injected intragingivally with Ad-C and Ad- Ch25h (1 × 10 9 PFU per 5 μl; n = 10). Scale bar, 100 μm. n indicates the number of biologically independent samples and mice per group. Values are presented as the mean ± SEM based on a two-tailed t -test ( c , j ) and one-way ANOVA with Tukey’s test ( b , e , f ). (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: For determination of cellular cholesterol levels, human GFs were treated with IL1β, TNFα, cholesterol synthesis inhibitors (triparanol (Cayman) and lovastatin (Enzo), or anti-CXCL16 blocking antibody (R&D Systems) in the presence of IL1β (2 ng ml −1 ), TNFα (50 ng ml −1 ) or cholesterol (200 μM).

Techniques: Microarray, Immunostaining, Infection, Staining, Injection, Virus, Two Tailed Test

Journal: Experimental & Molecular Medicine

Article Title: Disruption of cholesterol homeostasis triggers periodontal inflammation and alveolar bone loss

doi: 10.1038/s12276-023-01122-w

Figure Lengend Snippet: mRNA levels of CH25H , MMP1 , IL6 , IL8 , and PTGS2 in human GFs transfected with CH25H siRNA in the presence of IL1β (2 ng ml −1 ) ( a ) or TNFα (50 ng ml −1 ) ( b ) for 24 h ( n ≥ 3). Representative μCT ( c ) and H&E staining images ( d ) for BMD analysis and CEJ-ABC distance measurement of the maxillae of ligature-induced PD wild-type (WT) and CH25H knockout ( Ch25h −/− ) mice ( n ≥ 10). Scale bar, 100 μm. n indicates the number of biologically independent samples and mice per group. Values are presented as the mean ± SEM based on a two-tailed t -test ( c , d ) and one-way ANOVA with Tukey’s test ( a , b ). (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: For determination of cellular cholesterol levels, human GFs were treated with IL1β, TNFα, cholesterol synthesis inhibitors (triparanol (Cayman) and lovastatin (Enzo), or anti-CXCL16 blocking antibody (R&D Systems) in the presence of IL1β (2 ng ml −1 ), TNFα (50 ng ml −1 ) or cholesterol (200 μM).

Techniques: Transfection, Staining, Knock-Out, Two Tailed Test

Journal: Experimental & Molecular Medicine

Article Title: Disruption of cholesterol homeostasis triggers periodontal inflammation and alveolar bone loss

doi: 10.1038/s12276-023-01122-w

Figure Lengend Snippet: a Transcript levels of oxysterol-binding nuclear receptors in human GFs treated with IL1β (5 ng ml −1 ), TNFα (100 ng ml −1 ), cholesterol (200 μM), or 25-HC (20 μM) for 24 h ( n = 3). b RORα transcriptional activity in human GFs treated with IL1β (2 ng ml −1 ), TNFα (50 ng ml −1 ), cholesterol (100 μM), and 25-HC (20 μM) for 24 h or infected with Ad- CH25H (800 MOI; n = 4). Representative RORα immunostaining images of the gingiva of human patients with PD ( c ) and ligature-induced PD mice ( d ). Scale bar, 25 μm. mRNA ( e ) and protein ( f ) levels in human GFs infected with Ad- RORα for 48 h ( n ≥ 3). Representative RORα immunostaining ( g ), μCT, and H&E staining images ( h ) in mouse gingiva injected with an empty virus (Ad-C) or Ad- Rorα (1 × 10 9 PFU per 8 μl). Scale bar, 100 μm (left). Scale bar, 25 μm (right) ( g ). Scale bar, 100 μm ( h ). i Analysis of BMD and the CEJ-ABC distance in mouse maxillae injected intragingivally with Ad-C or Ad- Rorα (1 × 10 9 PFU per 8 μl; n = 10). n indicates the number of biologically independent samples and mice per group. Values are presented as the mean ± SEM based on a two-tailed t -test ( i ) and one-way ANOVA with Tukey’s test ( a , b , e ). (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: For determination of cellular cholesterol levels, human GFs were treated with IL1β, TNFα, cholesterol synthesis inhibitors (triparanol (Cayman) and lovastatin (Enzo), or anti-CXCL16 blocking antibody (R&D Systems) in the presence of IL1β (2 ng ml −1 ), TNFα (50 ng ml −1 ) or cholesterol (200 μM).

Techniques: Binding Assay, Activity Assay, Infection, Immunostaining, Staining, Injection, Virus, Two Tailed Test

Journal: Experimental & Molecular Medicine

Article Title: Disruption of cholesterol homeostasis triggers periodontal inflammation and alveolar bone loss

doi: 10.1038/s12276-023-01122-w

Figure Lengend Snippet: mRNA levels of human GFs treated with IL1β ( a ), TNFα ( b ), or cholesterol ( c ) for 24 h ( n ≥ 3) in the absence or presence of SR3335. Representative μCT and H&E staining images ( d ) of the analysis of BMD and the CEJ-ABC distance ( e ) of the maxillae of ligature-induced PD mice injected intragingivally with DMSO and SR3335 (15 mg kg −1 ; n ≥ 6). Scale bar, 100 μm. Representative μCT and H&E staining images ( f ) of the analysis of BMD and the CEJ-ABC distance ( g ) of the maxillae of mice fed a regular diet (RD) or high-cholesterol diet (HCD) and injected intragingivally with DMSO or SR3335 (15 mg kg −1 ; n ≥ 10). Scale bar, 100 μm. Values are presented as the mean ± SEM based on a two-tailed t -test ( e , g ) and one-way ANOVA with Tukey’s test ( a–c ). (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: For determination of cellular cholesterol levels, human GFs were treated with IL1β, TNFα, cholesterol synthesis inhibitors (triparanol (Cayman) and lovastatin (Enzo), or anti-CXCL16 blocking antibody (R&D Systems) in the presence of IL1β (2 ng ml −1 ), TNFα (50 ng ml −1 ) or cholesterol (200 μM).

Techniques: Staining, Injection, Two Tailed Test

Journal: PLoS ONE

Article Title: Multiple Cationic Amphiphiles Induce a Niemann-Pick C Phenotype and Inhibit Ebola Virus Entry and Infection

doi: 10.1371/journal.pone.0056265

Figure Lengend Snippet: SNB19 cells were pretreated with inhibitor for 1 hr, and EBOV VLP-GPΔ was then bound by spinfection at 4°C for 1 hr. Cells were then washed in media with inhibitor, incubated at 37°C for 3 hr (in inhibitor), and then processed for VLP entry as described in the section. Compounds were tested in multiple (n) experiments (each in triplicate) at either the highest concentration under which no toxicity was observed (for compounds that did not inhibit infection), or at the concentration that resulted in maximum inhibition of EBOV infection with minimal toxicity: SR 12813 (5 µM), (n = 4), colestolone (10 µM), (n = 5), alendronate (20 µM), (n = 3), BM 15766 (2 µM), (n = 4), amorolfine (6 µM), (n = 4), AY-9944 (5 µM), (n = 3), clomiphene (5 µM), (n = 9), Ro 48-8071 (5 µM, (n = 6), U18666A (5 µM), (n = 7), terconazole (10 µM), (n = 4), and triparanol (5 µM), (n = 3). Error bars represent standard error: * (P<2.96×10 −3 ) or ** (P<6.14×10 −5 ). Dashed line represents the observed threshold for entry inhibition needed to observe corresponding inhibition of live EBOV infection . As indicated in the key, colors denote classes of molecules.

Article Snippet: Chemicals were obtained from the following sources: 5-(N-Ethyl-N-isopropyl) amiloride (EIPA; CAS 1154-25-2), clomiphene citrate (CAS 50-41-9), triparanol (CAS 78-41-1), BM 15766 (CAS 86621-94-5), SR 12813 (CAS 126411-39-0), and Filipin (CAS 480-49-9) (Sigma-Aldrich); bafilomycin A1 (CAS 88899-55-2) (LC Laboratories); U18666A (CAS 3039-71-2) and E64d (CAS 88321-09-9) (EMD Biosciences; Ro 48-8071 (CAS 161582-11-2) (BIOMOL); AY-9944 (CAS 366-93-8) (TOCRIS); alendronate sodium (CAS 129318-43-0) (ABATRA); terconazole (CAS 67915-31-5) (LEIRAS); amorolfine hydrochloride (CAS 106614-68-0) (LKT); colestolone (CAS 50673-97-7)(Fisher Scientific).

Techniques: Incubation, Concentration Assay, Infection, Inhibition

Journal: PLoS ONE

Article Title: Multiple Cationic Amphiphiles Induce a Niemann-Pick C Phenotype and Inhibit Ebola Virus Entry and Infection

doi: 10.1371/journal.pone.0056265

Figure Lengend Snippet: Summary: Properties and effects of sterol synthesis pathway inhibitors on EBOV entry and infection and on cholesterol accumulation in LE/Lys.

Article Snippet: Chemicals were obtained from the following sources: 5-(N-Ethyl-N-isopropyl) amiloride (EIPA; CAS 1154-25-2), clomiphene citrate (CAS 50-41-9), triparanol (CAS 78-41-1), BM 15766 (CAS 86621-94-5), SR 12813 (CAS 126411-39-0), and Filipin (CAS 480-49-9) (Sigma-Aldrich); bafilomycin A1 (CAS 88899-55-2) (LC Laboratories); U18666A (CAS 3039-71-2) and E64d (CAS 88321-09-9) (EMD Biosciences; Ro 48-8071 (CAS 161582-11-2) (BIOMOL); AY-9944 (CAS 366-93-8) (TOCRIS); alendronate sodium (CAS 129318-43-0) (ABATRA); terconazole (CAS 67915-31-5) (LEIRAS); amorolfine hydrochloride (CAS 106614-68-0) (LKT); colestolone (CAS 50673-97-7)(Fisher Scientific).

Techniques: Infection