Journal: iScience

Article Title: PHLPP1 promotes neutral lipid accumulation through AMPK/ChREBP-dependent lipid uptake and fatty acid synthesis pathways

doi: 10.1016/j.isci.2022.103766

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal Anti-ABCG1 , Novus Biologicals , NB400-132.

Techniques: Recombinant, Clinical Proteomics, cDNA Synthesis, Plasmid Preparation, Software

Journal: Journal of Natural Products

Article Title: Leoligin, the Major Lignan from Edelweiss ( Leontopodium nivale subsp. alpinum ), Promotes Cholesterol Efflux from THP-1 Macrophages

doi: 10.1021/acs.jnatprod.6b00227

Figure Lengend Snippet: Leoligin increases ABCA1 and ABCG1, but not SR-B1 protein expression. THP-1 macrophages were treated with leoligin (LEO, 10 μM), pioglitazone (PIO, 10 μM), or solvent control (DMSO) for 24 h. Then, cells were lysed with NP40 buffer. A 20 μg amount of total protein was analyzed by Western blot using anti-ABCA1 (A), -ABCG1 (B), or -SR-B1 (C) antibodies. Band intensities of four independent experiments were quantified. The bar graphs represent mean ± SD, and the statistical evaluation was performed by one-way ANOVA with the Bonferroni post-test. * p < 0.05 versus solvent control (DMSO); ** p < 0.01 versus DMSO; *** p < 0.001 versus DMSO; n.s. not significant versus DMSO. For ABCA1 protein level, 95% CI of difference is −1.246 to −0.3587 (LEO, 10 μM) and −1.479 to −0.5911 (PIO, 10 μM); for ABCG1 protein level, 95% CI of difference is −0.6935 to −0.1084 (LEO, 10 μM) and −0.9088 to −0.3237 (PIO, 10 μM); for SR-B1 protein level, 95% CI of difference is −0.3437 to 0.3445 (LEO, 10 μM) and −0.4903 to 0.1979 (PIO, 10 μM).

Article Snippet: Primary antibodies against ABCA1, ABCG1, and SR-B1 were purchased from Novus Biologicals (Vienna, Austria).

Techniques: Expressing, Solvent, Control, Western Blot

Journal: Journal of Natural Products

Article Title: Leoligin, the Major Lignan from Edelweiss ( Leontopodium nivale subsp. alpinum ), Promotes Cholesterol Efflux from THP-1 Macrophages

doi: 10.1021/acs.jnatprod.6b00227

Figure Lengend Snippet: Leoligin increases the expression of ABCA1 (A) and ABCG1 (B) mRNA in THP-1 macrophages. THP-1 macrophages were treated with leoligin (LEO, 10 μM), pioglitazone (PIO, 10 μM), or solvent control (DMSO). After 24 h, total RNA was extracted followed by cDNA synthesis. qPCR was performed and quantified based on four independent experiments. The data are presented as mean ± SD, and the statistical evaluation was performed by one-way ANOVA analysis with the Bonferroni post-test. *** p < 0.001 versus solvent control (DMSO); n.s. not significant versus DMSO. For ABCA1 mRNA level, 95% CI of difference is −1.803 to −1.064 (LEO, 10 μM) and −2.301 to −1.562 (PIO, 10 μM); for ABCG1 mRNA level, 95% CI of difference is −3.726 to −2.134 (LEO, 10 μM) and −4.069 to −2.477 (PIO, 10 μM);.

Article Snippet: Primary antibodies against ABCA1, ABCG1, and SR-B1 were purchased from Novus Biologicals (Vienna, Austria).

Techniques: Expressing, Solvent, Control, cDNA Synthesis

Journal: Journal of Natural Products

Article Title: Leoligin, the Major Lignan from Edelweiss ( Leontopodium nivale subsp. alpinum ), Promotes Cholesterol Efflux from THP-1 Macrophages

doi: 10.1021/acs.jnatprod.6b00227

Figure Lengend Snippet: Leoligin does not prevent ABCA1 (A) and ABCG1 (B) mRNA degradation. Differentiated THP-1 macrophages were treated with 10 μM leoligin or vehicle (DMSO). After 24 h incubation, cells were treated with 5 μg/mL actinomycin D (Act D) and lysed at the indicated time points. Total RNA was extracted followed by cDNA synthesis. qPCR was performed and quantified based on four independent experiments. The data are presented as mean ± SD, and the statistical evaluation was performed by two-way ANOVA analysis with the Bonferroni post-test. ** p < 0.01 versus solvent control (DMSO); n.s. not significant versus DMSO. For ABCA1 protein, the 95% CI of difference between DMSO and leoligin at different time points is −19.25 to 19.25, 6.647 to 45.14, −0.3661 to 38.13, −8.693 to 29.80; for ABCG1 protein, the 95% CI of difference between DMSO and leoligin at different time points is −18.70 to 18.70, −10.45 to 26.94, −7.388 to 30.00, −17.90 to 19.49.

Article Snippet: Primary antibodies against ABCA1, ABCG1, and SR-B1 were purchased from Novus Biologicals (Vienna, Austria).

Techniques: Incubation, cDNA Synthesis, Solvent, Control

Journal: The FASEB Journal

Article Title: Dissociated sterol-based liver X receptor agonists as therapeutics for chronic inflammatory diseases

doi: 10.1096/fj.201600244R

Figure Lengend Snippet: Sterol-based LXR agonists attenuate DSS-induced colitis progression in mice. A) EC50 and fold change of LXR responding element (LXRE)-luciferase assay in THP1 cells, and liver microsome stability of DMHCA, MePipHCA, and T0901317. B) Mice were treated with vehicle (0.5% methyl cellulose/0.5% Tween 80) or compounds at doses as indicated for 4 d before administration of 3% DSS drinking water to induce colitis. After 5 d, 3% DSS water was replaced with regular drinking water. Mice were allowed to recover for another 6 d. Body weight of mice was monitored and recorded daily. Percentage of body weight change was calculated. No DSS: n = 3; other groups: n = 9. C) Representative hematoxylin and eosin staining of cross section of colon samples. D) Histology score of hematoxylin and eosin staining of colon samples. E) Gene expression of Il1b, Ccl2, Tnf, and Abcg1 of mouse colon samples. Results are presented as means ± sem. *P < 0.05, **P < 0.01.

Article Snippet: RNA was reverse transcribed into cDNA using qScript cDNA synthesis kit (Quanta BioSciences, Gaithersburg, MD, USA). qPCR was performed with PerfeCTa qPCR ToughMix (Quanta BioSciences) on 50 ng cDNA template/reaction using Taqman probes (Thermo Fisher Scientific Life Sciences ), including mouse genes Il1b (Mm00434228_m1), Ccl2 (Mm00441242_m1), Il6 (Mm00446190_m1), Tnf (Mm00443258_m1), Abcg1 (Mm00437390_m1), and human genes IL1B (Hs00174097_m1), chemokine (C-C motif) ligand 2 ( CCL2 ) (Hs00234140_m1), IL6 (Hs00985639_m1), IL8 (Hs00174103_m1), TNF (Hs01113624_g1), ATP-binding cassette transporter G1 ( ABCG1 ) (Hs00245154_m1).

Techniques: Luciferase, Staining, Gene Expression

Journal: The FASEB Journal

Article Title: Dissociated sterol-based liver X receptor agonists as therapeutics for chronic inflammatory diseases

doi: 10.1096/fj.201600244R

Figure Lengend Snippet: Sterol-based LXR agonists reduce inflammation in mouse LPMCs, mouse peritoneal cells, human colonic epithelial cells, and human PBMCs. A) LPMCs were isolated from mice treated with or without DSS water. Freshly isolated LPMCs were treated with 1 μM LXR agonists for 16 h before being collected for RNA expression analysis (upper panel), cytokine secretion analysis, and cell viability by CellTiter Glo assay (lower panel). B) Mouse peritoneal cells were treated with LXR agonists of different concentrations for 6 h before stimulation with LPS for 16 h, followed by HTRF TNF-α secretion assay. C) Mouse peritoneal cells were treated with 1 μM LXR agonists for 6 h before LPS stimulation for 1 h for gene expression analysis. D, E) SW480 cells were pretreated with control or compounds at concentrations as indicated for 16 h, before 10 ng/ml TNF-α (D) or 100 ng/ml LPS (E) stimulation. After 1 h, cells were collected and RNA was isolated. RT-qPCR was performed to determine the gene expression of CCL2, IL1B, IL6, IL8, TNF-α, ABCG1. F) Primary human PBMCs were treated with different LXR agonists for 6 h followed by LPS stimulation for 16 h. TNF-α secretion was assessed by HTRF assay and cell viability by CellTiter Glo assay. Results are presented as means ± sd.

Article Snippet: RNA was reverse transcribed into cDNA using qScript cDNA synthesis kit (Quanta BioSciences, Gaithersburg, MD, USA). qPCR was performed with PerfeCTa qPCR ToughMix (Quanta BioSciences) on 50 ng cDNA template/reaction using Taqman probes (Thermo Fisher Scientific Life Sciences ), including mouse genes Il1b (Mm00434228_m1), Ccl2 (Mm00441242_m1), Il6 (Mm00446190_m1), Tnf (Mm00443258_m1), Abcg1 (Mm00437390_m1), and human genes IL1B (Hs00174097_m1), chemokine (C-C motif) ligand 2 ( CCL2 ) (Hs00234140_m1), IL6 (Hs00985639_m1), IL8 (Hs00174103_m1), TNF (Hs01113624_g1), ATP-binding cassette transporter G1 ( ABCG1 ) (Hs00245154_m1).

Techniques: Isolation, RNA Expression, Glo Assay, Gene Expression, Control, Quantitative RT-PCR, HTRF Assay

Journal: The FASEB Journal

Article Title: Dissociated sterol-based liver X receptor agonists as therapeutics for chronic inflammatory diseases

doi: 10.1096/fj.201600244R

Figure Lengend Snippet: Sterol-based LXR agonists activate LXR in the brain and reduce inflammation induced by traumatic brain injury (TBI). A) Gene expression analysis of Abca1 and Abcg1 in brain tissues that were collected from mice treated with LXR agonists for 6 d (n = 6). B) Concentrations of LXR agonists in the plasma and brain from mice that were treated with a single dose of LXR agonists. C) LXR agonists activity profile in SHSY5Y cells. D) ABCA1 mRNA expression in SHSY5Y cells that were treated with 1 μM of different LXR agonists for various times. E) TBI experimental design. Compounds or vehicle were given by oral gavage once daily for 3 d before TBI induction. The final dose was given 15 min after injury induction. Tissues were collected 24 h postinjury (n = 5). F) Gene expression analysis of injured brain tissues. Results are presented as means ± sem (A, B, D), and as means ± sd (C). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: RNA was reverse transcribed into cDNA using qScript cDNA synthesis kit (Quanta BioSciences, Gaithersburg, MD, USA). qPCR was performed with PerfeCTa qPCR ToughMix (Quanta BioSciences) on 50 ng cDNA template/reaction using Taqman probes (Thermo Fisher Scientific Life Sciences ), including mouse genes Il1b (Mm00434228_m1), Ccl2 (Mm00441242_m1), Il6 (Mm00446190_m1), Tnf (Mm00443258_m1), Abcg1 (Mm00437390_m1), and human genes IL1B (Hs00174097_m1), chemokine (C-C motif) ligand 2 ( CCL2 ) (Hs00234140_m1), IL6 (Hs00985639_m1), IL8 (Hs00174103_m1), TNF (Hs01113624_g1), ATP-binding cassette transporter G1 ( ABCG1 ) (Hs00245154_m1).

Techniques: Gene Expression, Clinical Proteomics, Activity Assay, Expressing

Journal: Lipids in Health and Disease

Article Title: Sphingomyelin synthase overexpression increases cholesterol accumulation and decreases cholesterol secretion in liver cells

doi: 10.1186/1476-511X-10-46

Figure Lengend Snippet: The expression of ABCA1, ABCG1 and SR-BI . A . RT-PCR analysis; B . Western blot analysis. Values shown are means ± SD ( n = 3), * P < 0.05; ** P < 0.001 compared to control (Huh7-tTA).

Article Snippet: Proteins were electroblotted to a nitrocellulose membrane that had been incubated with respective antibodies: FLAG (CST), Apo A-I (Epitomics), ABCA1 (Boster), ABCG1 (Boster), or SR-BI (Epitomics).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control

Journal: Investigative Ophthalmology & Visual Science

Article Title: Cholesterol Accumulation Promotes Photoreceptor Senescence and Retinal Degeneration

doi: 10.1167/iovs.65.10.29

Figure Lengend Snippet: QPCR Probes

Article Snippet: Abcg1 , Mm01348250_m1.

Techniques:

Journal: Investigative Ophthalmology & Visual Science

Article Title: Cholesterol Accumulation Promotes Photoreceptor Senescence and Retinal Degeneration

doi: 10.1167/iovs.65.10.29

Figure Lengend Snippet: Characterization of cellular senescence in photoreceptor cells treated with cholesterol in vitro . mRNA expression of Abca1 , Abcg1 ( A ), and senescence markers ( p16 and p21 ) ( B ) in photoreceptor cell line 661W treated with cholesterol for 24 hours. ( C ) ISH ( p16 and p21 ) ( red ) of photoreceptor cell line treated with cholesterol. ( D ) mRNA expression of inflammatory cytokines ( Tnf-α , Cxcl12b , and Ccl2 ) in a photoreceptor cell line treated with cholesterol. ( E ) qPCR senescence array. ( F ) Immunofluorescence of photoreceptor cell line stained for Ki67 ( green ). Nuclei were stained with DAPI ( blue ). ( G ) The quantification of Ki67-positive cells. * P < 0.05; ** P < 0.01, *** P < 0.001, **** P < 0.0001, t -test for comparison between two groups, one-way ANOVA followed by Bonferroni correction for multiple comparisons.

Article Snippet: Abcg1 , Mm01348250_m1.

Techniques: In Vitro, Expressing, Immunofluorescence, Staining, Comparison