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antibodies against nr1d1  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc antibodies against nr1d1
    Antibodies Against Nr1d1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 101 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/nr1d1+antibody/Rev-Erbalpha+Rabbit+mAb/pm41163542-214-7-18
    Average 95 stars, based on 101 article reviews
    antibodies against nr1d1 - by Bioz Stars, 2026-10
    95/100 stars

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    Related Articles

    Incubation:

    Article Title: Core-Clock Genes Regulate Proliferation and Invasion via a Reciprocal Interplay with MACC1 in Colorectal Cancer Cells
    Article Snippet: For the protein–protein interaction assay, DynabeadsTM Protein G beads were used according to the manufacturer’s instructions (Invitrogen). .. In short, 1500 μg total protein was incubated with the NR1D1 antibody (2 μg, #13418, Cell Signaling, Danvers, MA, USA), the ARNTL antibody (2 μg ab93806, Abcam, Cambridge, UK) or an isotype control (ab172730, Abcam) overnight and pulled down using protein G magnetic beads. .. After elution, MACC1 (1/5000, HPA020081, Sigma-Aldrich, St. Louis, MO, USA) was detected by Western blotting.

    Article Title: Clock Gene Nr1d1 Alleviates Retinal Inflammation Through Repression of Hmga2 in Microglia
    Article Snippet: .. For immunoprecipitation, the 10% lysis sonicated chromatin was set as “Input”, 80% was applied in immunoprecipitation reactions with NR1D1 antibody (CST 13418) named “IP” while the remaining 10% was incubated with rabbit anti-IgG antibody (CST) as the negative control named “IgG”. ..

    Article Title: Alleviation of Limosilactobacillus reuteri in polycystic ovary syndrome protects against circadian dysrhythmia-induced dyslipidemia via capric acid and GALR1 signaling
    Article Snippet: Forty μg protein was loaded onto 10% SDS gel and then the protein was transferred to nitrocellulose membrane (Merck Millipore, Massachusetts, USA). .. The nonspecific binding sites of membrane were blocked and incubated with diluted GALR1 antibody (1:500; #47567, Signalway Antibody, Maryland, USA), GALR2 antibody (1:500; #26459-1-AP, Proteintech, Wuhan, China), NR1D1 antibody (1:1000, #13418 S, Cell Signaling Technology, Massachusetts, USA; 1:1000, #14506-1-AP, Proteintech), NR1D2 antibody (1:500; #13906-1-AP, Proteintech), SREBP1 antibody (1:1000; #41878, Signalway Antibody), LXRa antibody (1:1000; #ab176323, Abcam, Cambridge, UK), RXRa antibody (1:1000; #ab125001, Abcam), INSIG2 antibody (1:500; #24766-1, Proteintech), P-AKT antibody (1:1000; #4060, Cell Signaling Technology), T-AKT antibody (1:1000; #4691, Cell Signaling Technology), P-ERK antibody (1:1000; #4370, Cell Signaling Technology) and T-ERK antibody (1:1000; #4695, Cell Signaling Technology) at 4°C for overnight. .. Then the nitrocellulose membrane was incubated with diluted matched peroxidase-conjugated secondary antibodies (1:5000; Signalway Antibody) for 1 h at room temperature.

    Control:

    Article Title: Core-Clock Genes Regulate Proliferation and Invasion via a Reciprocal Interplay with MACC1 in Colorectal Cancer Cells
    Article Snippet: For the protein–protein interaction assay, DynabeadsTM Protein G beads were used according to the manufacturer’s instructions (Invitrogen). .. In short, 1500 μg total protein was incubated with the NR1D1 antibody (2 μg, #13418, Cell Signaling, Danvers, MA, USA), the ARNTL antibody (2 μg ab93806, Abcam, Cambridge, UK) or an isotype control (ab172730, Abcam) overnight and pulled down using protein G magnetic beads. .. After elution, MACC1 (1/5000, HPA020081, Sigma-Aldrich, St. Louis, MO, USA) was detected by Western blotting.

    Magnetic Beads:

    Article Title: Core-Clock Genes Regulate Proliferation and Invasion via a Reciprocal Interplay with MACC1 in Colorectal Cancer Cells
    Article Snippet: For the protein–protein interaction assay, DynabeadsTM Protein G beads were used according to the manufacturer’s instructions (Invitrogen). .. In short, 1500 μg total protein was incubated with the NR1D1 antibody (2 μg, #13418, Cell Signaling, Danvers, MA, USA), the ARNTL antibody (2 μg ab93806, Abcam, Cambridge, UK) or an isotype control (ab172730, Abcam) overnight and pulled down using protein G magnetic beads. .. After elution, MACC1 (1/5000, HPA020081, Sigma-Aldrich, St. Louis, MO, USA) was detected by Western blotting.

    Immunoprecipitation:

    Article Title: Clock Gene Nr1d1 Alleviates Retinal Inflammation Through Repression of Hmga2 in Microglia
    Article Snippet: .. For immunoprecipitation, the 10% lysis sonicated chromatin was set as “Input”, 80% was applied in immunoprecipitation reactions with NR1D1 antibody (CST 13418) named “IP” while the remaining 10% was incubated with rabbit anti-IgG antibody (CST) as the negative control named “IgG”. ..

    Lysis:

    Article Title: Clock Gene Nr1d1 Alleviates Retinal Inflammation Through Repression of Hmga2 in Microglia
    Article Snippet: .. For immunoprecipitation, the 10% lysis sonicated chromatin was set as “Input”, 80% was applied in immunoprecipitation reactions with NR1D1 antibody (CST 13418) named “IP” while the remaining 10% was incubated with rabbit anti-IgG antibody (CST) as the negative control named “IgG”. ..

    Sonication:

    Article Title: Clock Gene Nr1d1 Alleviates Retinal Inflammation Through Repression of Hmga2 in Microglia
    Article Snippet: .. For immunoprecipitation, the 10% lysis sonicated chromatin was set as “Input”, 80% was applied in immunoprecipitation reactions with NR1D1 antibody (CST 13418) named “IP” while the remaining 10% was incubated with rabbit anti-IgG antibody (CST) as the negative control named “IgG”. ..

    Negative Control:

    Article Title: Clock Gene Nr1d1 Alleviates Retinal Inflammation Through Repression of Hmga2 in Microglia
    Article Snippet: .. For immunoprecipitation, the 10% lysis sonicated chromatin was set as “Input”, 80% was applied in immunoprecipitation reactions with NR1D1 antibody (CST 13418) named “IP” while the remaining 10% was incubated with rabbit anti-IgG antibody (CST) as the negative control named “IgG”. ..

    Binding Assay:

    Article Title: Alleviation of Limosilactobacillus reuteri in polycystic ovary syndrome protects against circadian dysrhythmia-induced dyslipidemia via capric acid and GALR1 signaling
    Article Snippet: Forty μg protein was loaded onto 10% SDS gel and then the protein was transferred to nitrocellulose membrane (Merck Millipore, Massachusetts, USA). .. The nonspecific binding sites of membrane were blocked and incubated with diluted GALR1 antibody (1:500; #47567, Signalway Antibody, Maryland, USA), GALR2 antibody (1:500; #26459-1-AP, Proteintech, Wuhan, China), NR1D1 antibody (1:1000, #13418 S, Cell Signaling Technology, Massachusetts, USA; 1:1000, #14506-1-AP, Proteintech), NR1D2 antibody (1:500; #13906-1-AP, Proteintech), SREBP1 antibody (1:1000; #41878, Signalway Antibody), LXRa antibody (1:1000; #ab176323, Abcam, Cambridge, UK), RXRa antibody (1:1000; #ab125001, Abcam), INSIG2 antibody (1:500; #24766-1, Proteintech), P-AKT antibody (1:1000; #4060, Cell Signaling Technology), T-AKT antibody (1:1000; #4691, Cell Signaling Technology), P-ERK antibody (1:1000; #4370, Cell Signaling Technology) and T-ERK antibody (1:1000; #4695, Cell Signaling Technology) at 4°C for overnight. .. Then the nitrocellulose membrane was incubated with diluted matched peroxidase-conjugated secondary antibodies (1:5000; Signalway Antibody) for 1 h at room temperature.

    Membrane:

    Article Title: Alleviation of Limosilactobacillus reuteri in polycystic ovary syndrome protects against circadian dysrhythmia-induced dyslipidemia via capric acid and GALR1 signaling
    Article Snippet: Forty μg protein was loaded onto 10% SDS gel and then the protein was transferred to nitrocellulose membrane (Merck Millipore, Massachusetts, USA). .. The nonspecific binding sites of membrane were blocked and incubated with diluted GALR1 antibody (1:500; #47567, Signalway Antibody, Maryland, USA), GALR2 antibody (1:500; #26459-1-AP, Proteintech, Wuhan, China), NR1D1 antibody (1:1000, #13418 S, Cell Signaling Technology, Massachusetts, USA; 1:1000, #14506-1-AP, Proteintech), NR1D2 antibody (1:500; #13906-1-AP, Proteintech), SREBP1 antibody (1:1000; #41878, Signalway Antibody), LXRa antibody (1:1000; #ab176323, Abcam, Cambridge, UK), RXRa antibody (1:1000; #ab125001, Abcam), INSIG2 antibody (1:500; #24766-1, Proteintech), P-AKT antibody (1:1000; #4060, Cell Signaling Technology), T-AKT antibody (1:1000; #4691, Cell Signaling Technology), P-ERK antibody (1:1000; #4370, Cell Signaling Technology) and T-ERK antibody (1:1000; #4695, Cell Signaling Technology) at 4°C for overnight. .. Then the nitrocellulose membrane was incubated with diluted matched peroxidase-conjugated secondary antibodies (1:5000; Signalway Antibody) for 1 h at room temperature.



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    Image Search Results


    Chronic ethanol ingestion altered the oscillation patterns of key circadian signaling molecules in mouse lungs. Graphs summarize expression profiles of key circadian regulators in mouse lungs at 4 h intervals over 24 h for ethanol-fed (EtOH, red) and control-fed (CON, blue) groups. Panels show expressions of (A) Bmal1 , (B) Clock , (C) Rorα , and (D) Rev-erbα analyzed by qPCR. N = 6–9 lungs per group. The circadian oscillation patterns of the four genes from ethanol-fed mouse lungs significantly changed compared to lungs from control-fed mice. Data are presented as mean ± SE. * Indicates change with p < 0.05 compared to control-fed (CON) group at the same time point.

    Journal: Frontiers in Medicine

    Article Title: Downregulation of RORα by alcohol promotes TGFβ and α-SMA expression in mouse lung fibroblasts

    doi: 10.3389/fmed.2026.1719787

    Figure Lengend Snippet: Chronic ethanol ingestion altered the oscillation patterns of key circadian signaling molecules in mouse lungs. Graphs summarize expression profiles of key circadian regulators in mouse lungs at 4 h intervals over 24 h for ethanol-fed (EtOH, red) and control-fed (CON, blue) groups. Panels show expressions of (A) Bmal1 , (B) Clock , (C) Rorα , and (D) Rev-erbα analyzed by qPCR. N = 6–9 lungs per group. The circadian oscillation patterns of the four genes from ethanol-fed mouse lungs significantly changed compared to lungs from control-fed mice. Data are presented as mean ± SE. * Indicates change with p < 0.05 compared to control-fed (CON) group at the same time point.

    Article Snippet: The membranes were blocked with 5% milk/TBS with 0.1% Tween 20 then incubated with antibodies for α-SMA (ab5694 at 1:3000, Abcam Waltham, MA), TGFβ (555,052 at 1:500, BD Pharmingen), fibronectin (sc-9068 at 1:1000, Santa Cruz Biotechnology), BMAL1 (MA5-25133 at 1:500, Invitrogen), CLOCK (5,157 at 1:1000, Cell Signaling), RORα (82930-1-RR at 1:1000, Proteintech), REV-ERBα (14506-1-AP at 1:1000, Proteintech), Per1 (sc-398890 at 1:500, Santa Cruz Biotechnology), Per2 (PA5-100107 at 1:500, Invitrogen), or GAPDH (G9545 at 1:50,000, Sigma-Aldrich) respectively, then incubated with an appropriate secondary antibody and exposed to Clarity Western ECL substrate (Bio-Rad Laboratories).

    Techniques: Expressing, Control

    Chronic ethanol ingestion disrupts temporal protein expression of key circadian signaling molecules in the mouse lungs. Protein levels from mouse lungs collected at ZT0 and ZT12 were analyzed for (A) BMAL1 (75 kD), (B) CLOCK (100 kD), (C) RORα (60 kD), and (D) REV-ERBα (75 kD) and normalized with GAPDH (36 kD) levels in the same blot by western analysis. All data reported as fold-change compared to control-fed at ZT0 (CON) group. Representative western immunoblots are shown above the corresponding graphs. N = 6 per group. Data are presented as mean ± SE. # indicates change with p < 0.005 compared with ZT0 CON group and * indicates changes with p < 0.05 compared with ZT12 CON group.

    Journal: Frontiers in Medicine

    Article Title: Downregulation of RORα by alcohol promotes TGFβ and α-SMA expression in mouse lung fibroblasts

    doi: 10.3389/fmed.2026.1719787

    Figure Lengend Snippet: Chronic ethanol ingestion disrupts temporal protein expression of key circadian signaling molecules in the mouse lungs. Protein levels from mouse lungs collected at ZT0 and ZT12 were analyzed for (A) BMAL1 (75 kD), (B) CLOCK (100 kD), (C) RORα (60 kD), and (D) REV-ERBα (75 kD) and normalized with GAPDH (36 kD) levels in the same blot by western analysis. All data reported as fold-change compared to control-fed at ZT0 (CON) group. Representative western immunoblots are shown above the corresponding graphs. N = 6 per group. Data are presented as mean ± SE. # indicates change with p < 0.005 compared with ZT0 CON group and * indicates changes with p < 0.05 compared with ZT12 CON group.

    Article Snippet: The membranes were blocked with 5% milk/TBS with 0.1% Tween 20 then incubated with antibodies for α-SMA (ab5694 at 1:3000, Abcam Waltham, MA), TGFβ (555,052 at 1:500, BD Pharmingen), fibronectin (sc-9068 at 1:1000, Santa Cruz Biotechnology), BMAL1 (MA5-25133 at 1:500, Invitrogen), CLOCK (5,157 at 1:1000, Cell Signaling), RORα (82930-1-RR at 1:1000, Proteintech), REV-ERBα (14506-1-AP at 1:1000, Proteintech), Per1 (sc-398890 at 1:500, Santa Cruz Biotechnology), Per2 (PA5-100107 at 1:500, Invitrogen), or GAPDH (G9545 at 1:50,000, Sigma-Aldrich) respectively, then incubated with an appropriate secondary antibody and exposed to Clarity Western ECL substrate (Bio-Rad Laboratories).

    Techniques: Expressing, Western Blot, Control

    Ethanol exposure in vitro inhibited key molecules in the circadian signaling pathway. Gene expression levels from synchronized and unsynchronized murine primary lung fibroblasts (mPLFs) ± ethanol (EtOH, 60 mM; 24 h) were analyzed for: (A,B) Bmal1 , (C,D) Clock , (E,F) Rorα , and (G,H) Rev-erbα mRNA expression. Ethanol exposure significantly inhibited BMAL1, RORα, and Rev-erbα gene expressions while upregulating Clock gene expression. N = 6–10 per group. Data are presented as mean ± SE. * Indicates change with p < 0.05 compared to control (CON) group.

    Journal: Frontiers in Medicine

    Article Title: Downregulation of RORα by alcohol promotes TGFβ and α-SMA expression in mouse lung fibroblasts

    doi: 10.3389/fmed.2026.1719787

    Figure Lengend Snippet: Ethanol exposure in vitro inhibited key molecules in the circadian signaling pathway. Gene expression levels from synchronized and unsynchronized murine primary lung fibroblasts (mPLFs) ± ethanol (EtOH, 60 mM; 24 h) were analyzed for: (A,B) Bmal1 , (C,D) Clock , (E,F) Rorα , and (G,H) Rev-erbα mRNA expression. Ethanol exposure significantly inhibited BMAL1, RORα, and Rev-erbα gene expressions while upregulating Clock gene expression. N = 6–10 per group. Data are presented as mean ± SE. * Indicates change with p < 0.05 compared to control (CON) group.

    Article Snippet: The membranes were blocked with 5% milk/TBS with 0.1% Tween 20 then incubated with antibodies for α-SMA (ab5694 at 1:3000, Abcam Waltham, MA), TGFβ (555,052 at 1:500, BD Pharmingen), fibronectin (sc-9068 at 1:1000, Santa Cruz Biotechnology), BMAL1 (MA5-25133 at 1:500, Invitrogen), CLOCK (5,157 at 1:1000, Cell Signaling), RORα (82930-1-RR at 1:1000, Proteintech), REV-ERBα (14506-1-AP at 1:1000, Proteintech), Per1 (sc-398890 at 1:500, Santa Cruz Biotechnology), Per2 (PA5-100107 at 1:500, Invitrogen), or GAPDH (G9545 at 1:50,000, Sigma-Aldrich) respectively, then incubated with an appropriate secondary antibody and exposed to Clarity Western ECL substrate (Bio-Rad Laboratories).

    Techniques: In Vitro, Gene Expression, Expressing, Control

    Ethanol treatment inhibited core circadian signaling molecules in murine lung fibroblasts. Protein levels from synchronized and unsynchronized murine primary lung (mPLFs) ± ethanol (EtOH, 60 mM; 72 h) were analyzed for: (A,B) BMAL1 (75 kD), (C,D) CLOCK (100 kD), (E,F) RORα (60 kD), and (G,H) REV-ERBα (75 kD) were normalized to GAPDH (36 kD) levels from the same blot and reported as fold-change compared to the untreated (CON) group. Ethanol treatment significantly decreased the BAML1 and RORα protein levels in both synchronized and unsynchronized conditions, decreased REV-ERBα in the unsynchronized condition while it did not change the CLOCK protein expression in either condition. Representative western immunoblots are shown above the graphs. N = 8–15 per group. Data are presented as mean ± SE. * Indicates change with p < 0.05 compared to control (CON) group.

    Journal: Frontiers in Medicine

    Article Title: Downregulation of RORα by alcohol promotes TGFβ and α-SMA expression in mouse lung fibroblasts

    doi: 10.3389/fmed.2026.1719787

    Figure Lengend Snippet: Ethanol treatment inhibited core circadian signaling molecules in murine lung fibroblasts. Protein levels from synchronized and unsynchronized murine primary lung (mPLFs) ± ethanol (EtOH, 60 mM; 72 h) were analyzed for: (A,B) BMAL1 (75 kD), (C,D) CLOCK (100 kD), (E,F) RORα (60 kD), and (G,H) REV-ERBα (75 kD) were normalized to GAPDH (36 kD) levels from the same blot and reported as fold-change compared to the untreated (CON) group. Ethanol treatment significantly decreased the BAML1 and RORα protein levels in both synchronized and unsynchronized conditions, decreased REV-ERBα in the unsynchronized condition while it did not change the CLOCK protein expression in either condition. Representative western immunoblots are shown above the graphs. N = 8–15 per group. Data are presented as mean ± SE. * Indicates change with p < 0.05 compared to control (CON) group.

    Article Snippet: The membranes were blocked with 5% milk/TBS with 0.1% Tween 20 then incubated with antibodies for α-SMA (ab5694 at 1:3000, Abcam Waltham, MA), TGFβ (555,052 at 1:500, BD Pharmingen), fibronectin (sc-9068 at 1:1000, Santa Cruz Biotechnology), BMAL1 (MA5-25133 at 1:500, Invitrogen), CLOCK (5,157 at 1:1000, Cell Signaling), RORα (82930-1-RR at 1:1000, Proteintech), REV-ERBα (14506-1-AP at 1:1000, Proteintech), Per1 (sc-398890 at 1:500, Santa Cruz Biotechnology), Per2 (PA5-100107 at 1:500, Invitrogen), or GAPDH (G9545 at 1:50,000, Sigma-Aldrich) respectively, then incubated with an appropriate secondary antibody and exposed to Clarity Western ECL substrate (Bio-Rad Laboratories).

    Techniques: Expressing, Western Blot, Control

    Chronic ethanol ingestion altered the oscillation patterns of key circadian signaling molecules in mouse lungs. Graphs summarize expression profiles of key circadian regulators in mouse lungs at 4 h intervals over 24 h for ethanol-fed (EtOH, red) and control-fed (CON, blue) groups. Panels show expressions of (A) Bmal1 , (B) Clock , (C) Rorα , and (D) Rev-erbα analyzed by qPCR. N = 6–9 lungs per group. The circadian oscillation patterns of the four genes from ethanol-fed mouse lungs significantly changed compared to lungs from control-fed mice. Data are presented as mean ± SE. * Indicates change with p < 0.05 compared to control-fed (CON) group at the same time point.

    Journal: Frontiers in Medicine

    Article Title: Downregulation of RORα by alcohol promotes TGFβ and α-SMA expression in mouse lung fibroblasts

    doi: 10.3389/fmed.2026.1719787

    Figure Lengend Snippet: Chronic ethanol ingestion altered the oscillation patterns of key circadian signaling molecules in mouse lungs. Graphs summarize expression profiles of key circadian regulators in mouse lungs at 4 h intervals over 24 h for ethanol-fed (EtOH, red) and control-fed (CON, blue) groups. Panels show expressions of (A) Bmal1 , (B) Clock , (C) Rorα , and (D) Rev-erbα analyzed by qPCR. N = 6–9 lungs per group. The circadian oscillation patterns of the four genes from ethanol-fed mouse lungs significantly changed compared to lungs from control-fed mice. Data are presented as mean ± SE. * Indicates change with p < 0.05 compared to control-fed (CON) group at the same time point.

    Article Snippet: The membranes were blocked with 5% milk/TBS with 0.1% Tween 20 then incubated with antibodies for α-SMA (ab5694 at 1:3000, Abcam Waltham, MA), TGFβ (555,052 at 1:500, BD Pharmingen), fibronectin (sc-9068 at 1:1000, Santa Cruz Biotechnology), BMAL1 (MA5-25133 at 1:500, Invitrogen), CLOCK (5,157 at 1:1000, Cell Signaling), RORα (82930-1-RR at 1:1000, Proteintech), REV-ERBα (14506-1-AP at 1:1000, Proteintech), Per1 (sc-398890 at 1:500, Santa Cruz Biotechnology), Per2 (PA5-100107 at 1:500, Invitrogen), or GAPDH (G9545 at 1:50,000, Sigma-Aldrich) respectively, then incubated with an appropriate secondary antibody and exposed to Clarity Western ECL substrate (Bio-Rad Laboratories).

    Techniques: Expressing, Control

    Chronic ethanol ingestion disrupts temporal protein expression of key circadian signaling molecules in the mouse lungs. Protein levels from mouse lungs collected at ZT0 and ZT12 were analyzed for (A) BMAL1 (75 kD), (B) CLOCK (100 kD), (C) RORα (60 kD), and (D) REV-ERBα (75 kD) and normalized with GAPDH (36 kD) levels in the same blot by western analysis. All data reported as fold-change compared to control-fed at ZT0 (CON) group. Representative western immunoblots are shown above the corresponding graphs. N = 6 per group. Data are presented as mean ± SE. # indicates change with p < 0.005 compared with ZT0 CON group and * indicates changes with p < 0.05 compared with ZT12 CON group.

    Journal: Frontiers in Medicine

    Article Title: Downregulation of RORα by alcohol promotes TGFβ and α-SMA expression in mouse lung fibroblasts

    doi: 10.3389/fmed.2026.1719787

    Figure Lengend Snippet: Chronic ethanol ingestion disrupts temporal protein expression of key circadian signaling molecules in the mouse lungs. Protein levels from mouse lungs collected at ZT0 and ZT12 were analyzed for (A) BMAL1 (75 kD), (B) CLOCK (100 kD), (C) RORα (60 kD), and (D) REV-ERBα (75 kD) and normalized with GAPDH (36 kD) levels in the same blot by western analysis. All data reported as fold-change compared to control-fed at ZT0 (CON) group. Representative western immunoblots are shown above the corresponding graphs. N = 6 per group. Data are presented as mean ± SE. # indicates change with p < 0.005 compared with ZT0 CON group and * indicates changes with p < 0.05 compared with ZT12 CON group.

    Article Snippet: The membranes were blocked with 5% milk/TBS with 0.1% Tween 20 then incubated with antibodies for α-SMA (ab5694 at 1:3000, Abcam Waltham, MA), TGFβ (555,052 at 1:500, BD Pharmingen), fibronectin (sc-9068 at 1:1000, Santa Cruz Biotechnology), BMAL1 (MA5-25133 at 1:500, Invitrogen), CLOCK (5,157 at 1:1000, Cell Signaling), RORα (82930-1-RR at 1:1000, Proteintech), REV-ERBα (14506-1-AP at 1:1000, Proteintech), Per1 (sc-398890 at 1:500, Santa Cruz Biotechnology), Per2 (PA5-100107 at 1:500, Invitrogen), or GAPDH (G9545 at 1:50,000, Sigma-Aldrich) respectively, then incubated with an appropriate secondary antibody and exposed to Clarity Western ECL substrate (Bio-Rad Laboratories).

    Techniques: Expressing, Western Blot, Control

    Ethanol exposure in vitro inhibited key molecules in the circadian signaling pathway. Gene expression levels from synchronized and unsynchronized murine primary lung fibroblasts (mPLFs) ± ethanol (EtOH, 60 mM; 24 h) were analyzed for: (A,B) Bmal1 , (C,D) Clock , (E,F) Rorα , and (G,H) Rev-erbα mRNA expression. Ethanol exposure significantly inhibited BMAL1, RORα, and Rev-erbα gene expressions while upregulating Clock gene expression. N = 6–10 per group. Data are presented as mean ± SE. * Indicates change with p < 0.05 compared to control (CON) group.

    Journal: Frontiers in Medicine

    Article Title: Downregulation of RORα by alcohol promotes TGFβ and α-SMA expression in mouse lung fibroblasts

    doi: 10.3389/fmed.2026.1719787

    Figure Lengend Snippet: Ethanol exposure in vitro inhibited key molecules in the circadian signaling pathway. Gene expression levels from synchronized and unsynchronized murine primary lung fibroblasts (mPLFs) ± ethanol (EtOH, 60 mM; 24 h) were analyzed for: (A,B) Bmal1 , (C,D) Clock , (E,F) Rorα , and (G,H) Rev-erbα mRNA expression. Ethanol exposure significantly inhibited BMAL1, RORα, and Rev-erbα gene expressions while upregulating Clock gene expression. N = 6–10 per group. Data are presented as mean ± SE. * Indicates change with p < 0.05 compared to control (CON) group.

    Article Snippet: The membranes were blocked with 5% milk/TBS with 0.1% Tween 20 then incubated with antibodies for α-SMA (ab5694 at 1:3000, Abcam Waltham, MA), TGFβ (555,052 at 1:500, BD Pharmingen), fibronectin (sc-9068 at 1:1000, Santa Cruz Biotechnology), BMAL1 (MA5-25133 at 1:500, Invitrogen), CLOCK (5,157 at 1:1000, Cell Signaling), RORα (82930-1-RR at 1:1000, Proteintech), REV-ERBα (14506-1-AP at 1:1000, Proteintech), Per1 (sc-398890 at 1:500, Santa Cruz Biotechnology), Per2 (PA5-100107 at 1:500, Invitrogen), or GAPDH (G9545 at 1:50,000, Sigma-Aldrich) respectively, then incubated with an appropriate secondary antibody and exposed to Clarity Western ECL substrate (Bio-Rad Laboratories).

    Techniques: In Vitro, Gene Expression, Expressing, Control

    Ethanol treatment inhibited core circadian signaling molecules in murine lung fibroblasts. Protein levels from synchronized and unsynchronized murine primary lung (mPLFs) ± ethanol (EtOH, 60 mM; 72 h) were analyzed for: (A,B) BMAL1 (75 kD), (C,D) CLOCK (100 kD), (E,F) RORα (60 kD), and (G,H) REV-ERBα (75 kD) were normalized to GAPDH (36 kD) levels from the same blot and reported as fold-change compared to the untreated (CON) group. Ethanol treatment significantly decreased the BAML1 and RORα protein levels in both synchronized and unsynchronized conditions, decreased REV-ERBα in the unsynchronized condition while it did not change the CLOCK protein expression in either condition. Representative western immunoblots are shown above the graphs. N = 8–15 per group. Data are presented as mean ± SE. * Indicates change with p < 0.05 compared to control (CON) group.

    Journal: Frontiers in Medicine

    Article Title: Downregulation of RORα by alcohol promotes TGFβ and α-SMA expression in mouse lung fibroblasts

    doi: 10.3389/fmed.2026.1719787

    Figure Lengend Snippet: Ethanol treatment inhibited core circadian signaling molecules in murine lung fibroblasts. Protein levels from synchronized and unsynchronized murine primary lung (mPLFs) ± ethanol (EtOH, 60 mM; 72 h) were analyzed for: (A,B) BMAL1 (75 kD), (C,D) CLOCK (100 kD), (E,F) RORα (60 kD), and (G,H) REV-ERBα (75 kD) were normalized to GAPDH (36 kD) levels from the same blot and reported as fold-change compared to the untreated (CON) group. Ethanol treatment significantly decreased the BAML1 and RORα protein levels in both synchronized and unsynchronized conditions, decreased REV-ERBα in the unsynchronized condition while it did not change the CLOCK protein expression in either condition. Representative western immunoblots are shown above the graphs. N = 8–15 per group. Data are presented as mean ± SE. * Indicates change with p < 0.05 compared to control (CON) group.

    Article Snippet: The membranes were blocked with 5% milk/TBS with 0.1% Tween 20 then incubated with antibodies for α-SMA (ab5694 at 1:3000, Abcam Waltham, MA), TGFβ (555,052 at 1:500, BD Pharmingen), fibronectin (sc-9068 at 1:1000, Santa Cruz Biotechnology), BMAL1 (MA5-25133 at 1:500, Invitrogen), CLOCK (5,157 at 1:1000, Cell Signaling), RORα (82930-1-RR at 1:1000, Proteintech), REV-ERBα (14506-1-AP at 1:1000, Proteintech), Per1 (sc-398890 at 1:500, Santa Cruz Biotechnology), Per2 (PA5-100107 at 1:500, Invitrogen), or GAPDH (G9545 at 1:50,000, Sigma-Aldrich) respectively, then incubated with an appropriate secondary antibody and exposed to Clarity Western ECL substrate (Bio-Rad Laboratories).

    Techniques: Expressing, Western Blot, Control