Review

vdac d73d12 antibody (Cell Signaling Technology Inc)

Name: vdac d73d12 antibody
Brand: Cell Signaling Technology Inc
Rating: 90 (1 reviews)

Cell Signaling Technology Inc is a verified supplier

Cell Signaling Technology Inc manufactures this product

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Cell Signaling Technology Inc vdac d73d12 antibody
$(a) Schematic illustration of NAD + biosynthetic pathways. In the de novo pathway the essential amino acid tryptophan (TRP) from the diet is utilized to produce NAD + via several reactions (dashed arrow). The salvage pathway recycles NAM, which is generated as a by-product of NAD + -dependent enzymes such as SIRT3; PA=Phthalic Acid. (b-h) BMMs were isolated from young female C57BL/6 mice and cultured with M-CSF and RANKL, in the presence or absence of FK866 or PA for 24 h. (b) NAD + levels in whole cells. (c) Representative images and quantification of TRAP-positive osteoclasts after 5 days. (d) NAD + levels in mitochondrial-enriched fractions. (e) SIRT3 activity. (f) Oxygen Consumption Rate. (g) ATP levels. (h) Apoptosis by caspase-3 activity. (i-j) Human BMMs were cultured with M-CSF and RANKL in the presence or absence of FK866, as above. (i) NAD + levels after 24 h and (j) representative images and quantification of TRAP-positive osteoclasts after 7 days. (k) mRNA levels of the indicated enzymes in BMMs cultured with M-CSF and RANKL the presence or absence of E 2 for the indicated timepoints. P values in grey indicate comparisons with time 0. P values in red indicate comparison of RANKL vs RANKL+E 2 within the same time point. (l) BMMs were isolated from young female C57BL/6 mice and cultured with M-CSF and RANKL the presence or absence of E 2 for 24 hours, representative Western blot images and quantification of the indicated proteins in mitochondrial enriched fractions; β-actin in cytosol and <t>VDAC</t> in mitochondria indicate efficacy of cellular compartment isolation. (m) mRNA levels in BMMs transduced with Nmnat1 <t>and</t> <t>Nmnat3</t> sh or control sh lentivirus particles. (n) NAD + levels after 24 h and (o) representative images and quantification of TRAP-positive osteoclasts after 5 days. Scale bar 500 µm. Line and error bars represent mean ± SD. P values using two-tailed Student’s t-test, 1-way ANOVA followed by Dunnett’s multiple comparisons test, or 2-way ANOVA followed by Šídák’s multiple comparisons test.$
Vdac D73d12 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/vdac d73d12 antibody/product/Cell Signaling Technology Inc
Average 90 stars, based on 1 article reviews

vdac d73d12 antibody - by Bioz Stars, 2026-03

90/100 stars

Images

1) Product Images from "Estrogens protect bone mass by inhibiting NAD + metabolism in osteoclasts"

Article Title: Estrogens protect bone mass by inhibiting NAD + metabolism in osteoclasts

Journal: bioRxiv

doi: 10.1101/2025.07.11.664289

$(a) Schematic illustration of NAD + biosynthetic pathways. In the de novo pathway the essential amino acid tryptophan (TRP) from the diet is utilized to produce NAD + via several reactions (dashed arrow). The salvage pathway recycles NAM, which is generated as a by-product of NAD + -dependent enzymes such as SIRT3; PA=Phthalic Acid. (b-h) BMMs were isolated from young female C57BL/6 mice and cultured with M-CSF and RANKL, in the presence or absence of FK866 or PA for 24 h. (b) NAD + levels in whole cells. (c) Representative images and quantification of TRAP-positive osteoclasts after 5 days. (d) NAD + levels in mitochondrial-enriched fractions. (e) SIRT3 activity. (f) Oxygen Consumption Rate. (g) ATP levels. (h) Apoptosis by caspase-3 activity. (i-j) Human BMMs were cultured with M-CSF and RANKL in the presence or absence of FK866, as above. (i) NAD + levels after 24 h and (j) representative images and quantification of TRAP-positive osteoclasts after 7 days. (k) mRNA levels of the indicated enzymes in BMMs cultured with M-CSF and RANKL the presence or absence of E 2 for the indicated timepoints. P values in grey indicate comparisons with time 0. P values in red indicate comparison of RANKL vs RANKL+E 2 within the same time point. (l) BMMs were isolated from young female C57BL/6 mice and cultured with M-CSF and RANKL the presence or absence of E 2 for 24 hours, representative Western blot images and quantification of the indicated proteins in mitochondrial enriched fractions; β-actin in cytosol and VDAC in mitochondria indicate efficacy of cellular compartment isolation. (m) mRNA levels in BMMs transduced with Nmnat1 and Nmnat3 sh or control sh lentivirus particles. (n) NAD + levels after 24 h and (o) representative images and quantification of TRAP-positive osteoclasts after 5 days. Scale bar 500 µm. Line and error bars represent mean ± SD. P values using two-tailed Student’s t-test, 1-way ANOVA followed by Dunnett’s multiple comparisons test, or 2-way ANOVA followed by Šídák’s multiple comparisons test.$
Figure Legend Snippet: (a) Schematic illustration of NAD + biosynthetic pathways. In the de novo pathway the essential amino acid tryptophan (TRP) from the diet is utilized to produce NAD + via several reactions (dashed arrow). The salvage pathway recycles NAM, which is generated as a by-product of NAD + -dependent enzymes such as SIRT3; PA=Phthalic Acid. (b-h) BMMs were isolated from young female C57BL/6 mice and cultured with M-CSF and RANKL, in the presence or absence of FK866 or PA for 24 h. (b) NAD + levels in whole cells. (c) Representative images and quantification of TRAP-positive osteoclasts after 5 days. (d) NAD + levels in mitochondrial-enriched fractions. (e) SIRT3 activity. (f) Oxygen Consumption Rate. (g) ATP levels. (h) Apoptosis by caspase-3 activity. (i-j) Human BMMs were cultured with M-CSF and RANKL in the presence or absence of FK866, as above. (i) NAD + levels after 24 h and (j) representative images and quantification of TRAP-positive osteoclasts after 7 days. (k) mRNA levels of the indicated enzymes in BMMs cultured with M-CSF and RANKL the presence or absence of E 2 for the indicated timepoints. P values in grey indicate comparisons with time 0. P values in red indicate comparison of RANKL vs RANKL+E 2 within the same time point. (l) BMMs were isolated from young female C57BL/6 mice and cultured with M-CSF and RANKL the presence or absence of E 2 for 24 hours, representative Western blot images and quantification of the indicated proteins in mitochondrial enriched fractions; β-actin in cytosol and VDAC in mitochondria indicate efficacy of cellular compartment isolation. (m) mRNA levels in BMMs transduced with Nmnat1 and Nmnat3 sh or control sh lentivirus particles. (n) NAD + levels after 24 h and (o) representative images and quantification of TRAP-positive osteoclasts after 5 days. Scale bar 500 µm. Line and error bars represent mean ± SD. P values using two-tailed Student’s t-test, 1-way ANOVA followed by Dunnett’s multiple comparisons test, or 2-way ANOVA followed by Šídák’s multiple comparisons test.

Techniques Used: Generated, Isolation, Cell Culture, Activity Assay, Comparison, Western Blot, Transduction, Control, Two Tailed Test

(a) Chemical reaction catalyzed by Lb NOX. (b) Western blot for FLAG, β-actin, and VDAC from BMMs cultured with M-CSF expressing the FLAG-tagged cytosolic and mitochondrial (mito) Lb NOX. β-actin in cytosol and VDAC in mitochondria indicate efficacy of cellular compartment isolation. (c) NAD + /NADH redox ratio and (d) ATP levels in BMMs expressing Lb NOX or mito Lb NOX. (e-g) BMMs expressing Lb NOX or mito Lb NOX cultured with M-CSF and RANKL in the presence or absence of E 2 (10 -8 M) for 24 hours. (e) NAD + /NADH redox ratio. (f) SIRT3 activity. (g) Representative images and quantification of TRAP-positive osteoclasts after 5 days. Scale bar 500 µm. Line and error bars represent mean ± SD. P values by 1-way ANOVA followed by Dunnett’s multiple comparisons test or 2-way ANOVA followed by Šídák’s multiple comparisons test.

Figure Legend Snippet: (a) Chemical reaction catalyzed by Lb NOX. (b) Western blot for FLAG, β-actin, and VDAC from BMMs cultured with M-CSF expressing the FLAG-tagged cytosolic and mitochondrial (mito) Lb NOX. β-actin in cytosol and VDAC in mitochondria indicate efficacy of cellular compartment isolation. (c) NAD + /NADH redox ratio and (d) ATP levels in BMMs expressing Lb NOX or mito Lb NOX. (e-g) BMMs expressing Lb NOX or mito Lb NOX cultured with M-CSF and RANKL in the presence or absence of E 2 (10 -8 M) for 24 hours. (e) NAD + /NADH redox ratio. (f) SIRT3 activity. (g) Representative images and quantification of TRAP-positive osteoclasts after 5 days. Scale bar 500 µm. Line and error bars represent mean ± SD. P values by 1-way ANOVA followed by Dunnett’s multiple comparisons test or 2-way ANOVA followed by Šídák’s multiple comparisons test.

Techniques Used: Western Blot, Cell Culture, Expressing, Isolation, Activity Assay

Image Search Results

A. Confocal microscopy (left) showing mitochondrial calcium level (Rhod-2 fluorescence) in cultured (myo)fibroblasts from normal RV, cRV and dRV. Quantification values (right) are expressed as mean ± SEM; n = 50 cells from 3 animals for each group. **P < 0.01; ****P < 0.0001. B. PDH activity, measured by an NADH-based enzymatic activity ElLISA, in (myo)fibroblasts from normal RV (CTRL), cRV and dRV. Quantification values (right) are expressed as mean ± SEM; n = 50 cells from 3 experiments. n = 4 animals for each group. *P < 0.05; **P < 0.01. C. Confocal microscope (left) shows the PRMT1 (i.e. arginine methyltransferase) level in (myo)fibroblasts (vimentin-positive cells) from right ventricle frozen sections of Control, cRV and dRV rats. Quantification values (right) are expressed as mean ± SEM; n = 50 cells from 3 animals for each group. ****P < 0.0001. D. Confocal microscopy (left) shows the AMDA level (i.e. arginine methylation level) in (myo)fibroblasts (vimentin-positive cells) from Control, cRV and dRV tissues. Quantification values (right) are expressed as mean ± SEM; n = 50 cells from 3 animals for each group. *P < 0.05; **P < 0.01; ****P < 0.0001. E. Confocal microscopy (left) shows the cytoplasmic PRMT1 level in cultured cardiac (myo)fibroblasts from normal RV, cRV and dRVs. Quantification values (right) are expressed as mean ± SEM; n = 50 cells from 3 experiments. *P < 0.05; **P < 0.01; ****P < 0.0001. F. Confocal microscopy (left) shows the cytoplasmic ADMA levels in cultured (myo)fibroblasts from normal RV, cRV and dRVs. Quantification values (right) are expressed as mean ± SEM; n = 50 cells from 3 experiments. ***P < 0.001. G. Immunoblot (left) of PRMT1, ADMA, MICU1, VDAC1/2 and β-Actin in (myo)fibroblasts from normal RV, cRV and dRV of rats. Quantification values (right) are expressed as mean ± SEM; n = 4 animals for each group. *P < 0.05. H. MICU1 Immunoprecipitation (above) on isolated mitochondria from (myo)fibroblasts shows the amount of MICU1 and ADMA. MICU1 methylation is shown as ADMA/MICU1. Quantification values (below) are expressed as mean ± SEM; n = 4 animals for each group. A, C, D, E, F comparisons were made using one-way ANOVA followed by Bonferroni post hoc analysis; B, G, H were made using Kruskal–Wallis tests followed by pairwise Mann–Whitney U tests. CTRL , control (normal right ventricle, without monocrotaline injection); cRV , compensated right ventricle; dRV , decompensated right ventricle; cFB, cardiac fibroblasts; cMFB, cardiac myofibroblasts; c(M)FB , cardiac (myo)fibroblasts; PDH , pyruvate dehydrogenase; PRMT1 , Protein Arginine Methyltransferase 1; ADMA , Asymmetric dimethylarginine; DAPI , 4′,6-diamidino-2-phenylindole (labels the nucleus); MICU1 , Mitochondrial Calcium Uptake 1; VDAC , Voltage-Dependent Anion Channels; IP , Immunoprecipitation.

Journal: bioRxiv

Article Title: A critical contribution of cardiac myofibroblasts in right ventricular failure and the role of UCP2 SNPs in the predisposition to RV decompensation in pulmonary arterial hypertension

doi: 10.1101/2025.07.25.666908

Figure Lengend Snippet: A. Confocal microscopy (left) showing mitochondrial calcium level (Rhod-2 fluorescence) in cultured (myo)fibroblasts from normal RV, cRV and dRV. Quantification values (right) are expressed as mean ± SEM; n = 50 cells from 3 animals for each group. **P < 0.01; ****P < 0.0001. B. PDH activity, measured by an NADH-based enzymatic activity ElLISA, in (myo)fibroblasts from normal RV (CTRL), cRV and dRV. Quantification values (right) are expressed as mean ± SEM; n = 50 cells from 3 experiments. n = 4 animals for each group. *P < 0.05; **P < 0.01. C. Confocal microscope (left) shows the PRMT1 (i.e. arginine methyltransferase) level in (myo)fibroblasts (vimentin-positive cells) from right ventricle frozen sections of Control, cRV and dRV rats. Quantification values (right) are expressed as mean ± SEM; n = 50 cells from 3 animals for each group. ****P < 0.0001. D. Confocal microscopy (left) shows the AMDA level (i.e. arginine methylation level) in (myo)fibroblasts (vimentin-positive cells) from Control, cRV and dRV tissues. Quantification values (right) are expressed as mean ± SEM; n = 50 cells from 3 animals for each group. *P < 0.05; **P < 0.01; ****P < 0.0001. E. Confocal microscopy (left) shows the cytoplasmic PRMT1 level in cultured cardiac (myo)fibroblasts from normal RV, cRV and dRVs. Quantification values (right) are expressed as mean ± SEM; n = 50 cells from 3 experiments. *P < 0.05; **P < 0.01; ****P < 0.0001. F. Confocal microscopy (left) shows the cytoplasmic ADMA levels in cultured (myo)fibroblasts from normal RV, cRV and dRVs. Quantification values (right) are expressed as mean ± SEM; n = 50 cells from 3 experiments. ***P < 0.001. G. Immunoblot (left) of PRMT1, ADMA, MICU1, VDAC1/2 and β-Actin in (myo)fibroblasts from normal RV, cRV and dRV of rats. Quantification values (right) are expressed as mean ± SEM; n = 4 animals for each group. *P < 0.05. H. MICU1 Immunoprecipitation (above) on isolated mitochondria from (myo)fibroblasts shows the amount of MICU1 and ADMA. MICU1 methylation is shown as ADMA/MICU1. Quantification values (below) are expressed as mean ± SEM; n = 4 animals for each group. A, C, D, E, F comparisons were made using one-way ANOVA followed by Bonferroni post hoc analysis; B, G, H were made using Kruskal–Wallis tests followed by pairwise Mann–Whitney U tests. CTRL , control (normal right ventricle, without monocrotaline injection); cRV , compensated right ventricle; dRV , decompensated right ventricle; cFB, cardiac fibroblasts; cMFB, cardiac myofibroblasts; c(M)FB , cardiac (myo)fibroblasts; PDH , pyruvate dehydrogenase; PRMT1 , Protein Arginine Methyltransferase 1; ADMA , Asymmetric dimethylarginine; DAPI , 4′,6-diamidino-2-phenylindole (labels the nucleus); MICU1 , Mitochondrial Calcium Uptake 1; VDAC , Voltage-Dependent Anion Channels; IP , Immunoprecipitation.

Article Snippet: Antibodies and dilutions: PRMT1 (Cell Signaling, 2449S) 1:1000, ADMA (Cell Signaling, 13522S) 1:1000, MICU1 (Sigma Aldrich, HPA037480) 1:1000, MCU (Cell Signaling, 14997S) 1:1000, UCP2 (Cell Signaling, 89326S) 1:1000, VDAC 1⁄2 (Proteintech, 10866-1-AP) and β-Actin (Santa Cruz sc81178) 1:4,000.

Techniques: Confocal Microscopy, Fluorescence, Cell Culture, Activity Assay, Microscopy, Control, Methylation, Western Blot, Immunoprecipitation, Isolation, MANN-WHITNEY, Injection

Immunoblot shows the amount of PRMT1, ADMA, MICU1, VDAC1/2 and β-Actin in cardiomyocytes from normal RV, cRV and dRVs. Quantification values are expressed as mean ± SEM; n = 4 animals for each group. Comparisons were made using Kruskal–Wallis tests. CTRL , control (normal right ventricle, without monocrotaline injection); cRV , compensated right ventricle; dRV , decompensated right ventricle; PRMT1 , Protein Arginine Methyltransferase 1; ADMA , Asymmetric dimethylarginine; MICU1 , Mitochondrial Calcium Uptake 1; VDAC , Voltage-Dependent Anion Channels; CM , cardiomyocytes.

Journal: bioRxiv

doi: 10.1101/2025.07.25.666908

Figure Lengend Snippet: Immunoblot shows the amount of PRMT1, ADMA, MICU1, VDAC1/2 and β-Actin in cardiomyocytes from normal RV, cRV and dRVs. Quantification values are expressed as mean ± SEM; n = 4 animals for each group. Comparisons were made using Kruskal–Wallis tests. CTRL , control (normal right ventricle, without monocrotaline injection); cRV , compensated right ventricle; dRV , decompensated right ventricle; PRMT1 , Protein Arginine Methyltransferase 1; ADMA , Asymmetric dimethylarginine; MICU1 , Mitochondrial Calcium Uptake 1; VDAC , Voltage-Dependent Anion Channels; CM , cardiomyocytes.

Techniques: Western Blot, Control, Injection

A. qPCR shows the UCP2 RNA level in human RV (from cohort 1and 3) genotyped with WT, UCP2 HET and HOM SNP. Scatter dot plots represent individual values. Quantification values are expressed as mean ± SEM. ***P < 0.001. B. Western blot (left) shows the UCP2 protein level in human RV genotyped with WT, UCP2 HET and HOM SNP. Scatter dot plots represent individual values. Quantification values (right) are expressed as mean ± SEM; *P < 0.05. C. UCP2 genotyping and TAPSE in patients with PAH (mPAP>20mmHg from cohort 1). Patients were genotyped for UCP2 and categorized into WT (n=6), UCP2 HET SNP (n=5) and UCP2 HOM SNP (n=4). Scatter dot plots represent individual values. Quantification values (right) are expressed as mean ± SEM *P < 0.05; **P < 0.01. D. UCP2 genotyping and echocardiography show the UCP2 genotype and Cardiac Index in patients with PAH (mPAP>20mmHg from cohort 1), sub-grouped by UCP2 genotype: WT (n=8), UCP2 HET SNP (n=6) and UCP2 HOM SNP (n=4). Scatter dot plots represent individual values. Quantification values (right) are expressed as mean ± SEM. E. UCP2 genotyping and echocardiography show the UCP2 genotype (blood) and Cardiac index in patients with PAH (mPAP>20mmHg from cohort 2). Patients were genotyped for UCP2 and categorized into WT (n=10), UCP2 HET SNP (n=6) and UCP2 HOM SNP (n=5). Scatter dot plots represent individual values. Quantification values ( right ) are expressed as mean ± SEM. *P < 0.05; **P < 0.01. F. UCP2 genotyping and echocardiography show the UCP2 genotype and Cardiac Index in patients with PAH (mPAP>20mmHg from cohort 2). Patients were genotyped for UCP2 and categorized into WT (n=10), UCP2 HET SNP (n=6) and UCP2 HOM SNP (n=5). Scatter dot plots represent individual values. Quantification values ( right ) are expressed as mean ± SEM. G. UCP2 genotyping and echocardiography show the UCP2 genotype and TAPSE in patients with PAH (mPAP>20mmHg from cohort 1and 2). Patients were genotyped for UCP2 and categorized into WT (n=16), UCP2 HET SNP (n=11) and UCP2 HOM SNP (n=9). Scatter dot plots represent individual values. Quantification values (right) are expressed as mean ± SEM *P < 0.05; **P < 0.01; ***P < 0.001. H. UCP2 genotyping and echocardiography show the UCP2 genotype and Cardiac index in patients with PAH (mPAP>20mmHg from cohort 1and 2). Patients were genotyped for UCP2 and categorized into WT (n=18), UCP2 HET SNP (n=12) and UCP2 HOM SNP (n=9). Scatter dot plots represent individual values. Quantification values (right) are expressed as mean ± SEM. *P < 0.05. I. UCP2 genotyping shows the ratio of patients with WT, UCP2-HET SNP and UCP2-HOM SNP in cRV vs dRV in patients with PAH (Cohort 1+2). J. UCP2 genotype and TAPSE in patients with sPHT (mPAP>20 mmHg from Cohort 3). Patients were genotyped for UCP2 and categorized into WT (n=6), UCP2 HET SNP (n=8) and UCP2 HOM SNP (n=3). Scatter dot plots represent individual values. Quantification values ( right ) are expressed as mean ± SEM. K. UCP2 genotype and TAPSE in patients with with sPHT (mPAP>20 mmHg from Cohort 3). Patients were genotyped for UCP2 and categorized into WT (n=8), UCP2 HET SNP (n=9) and UCP2 HOM SNP (n=4). Scatter dot plots represent individual values. Quantification values ( right ) are expressed as mean ± SEM. L. Spearman correlation test shows lack of orrelation between UCP2 RNA level and TAPSE in patients with sPHT (mPAP>20 mmHg from Cohort 3). Scatter dot plots show individual values. These comparisons were made using Kruskal–Wallis tests followed by pairwise Mann–Whitney U tests. SNP, Single Nucleotide Polymorphism; UCP2 , uncoupling protein 2; VDAC , Voltage-Dependent Anion Channels; WT, wild type; UCP2 HET, heterozygous UCP2 SNP; UCP2 HOM, homozygous UCP2 SNP; TAPSE, Tricuspid Annular Plane Systolic Excursion; mPAP: mean PA pressure; PAH, pulmonary arterial hypertension; s PHT : Secondary pulmonary hypertension; nRV, normal right ventricle; cRV, compensated right ventricle; dRV , decompensated right ventricle; cFB, cardiac fibroblasts; cMFB, cardiac myofibroblasts.

Journal: bioRxiv

doi: 10.1101/2025.07.25.666908

Figure Lengend Snippet: A. qPCR shows the UCP2 RNA level in human RV (from cohort 1and 3) genotyped with WT, UCP2 HET and HOM SNP. Scatter dot plots represent individual values. Quantification values are expressed as mean ± SEM. ***P < 0.001. B. Western blot (left) shows the UCP2 protein level in human RV genotyped with WT, UCP2 HET and HOM SNP. Scatter dot plots represent individual values. Quantification values (right) are expressed as mean ± SEM; *P < 0.05. C. UCP2 genotyping and TAPSE in patients with PAH (mPAP>20mmHg from cohort 1). Patients were genotyped for UCP2 and categorized into WT (n=6), UCP2 HET SNP (n=5) and UCP2 HOM SNP (n=4). Scatter dot plots represent individual values. Quantification values (right) are expressed as mean ± SEM *P < 0.05; **P < 0.01. D. UCP2 genotyping and echocardiography show the UCP2 genotype and Cardiac Index in patients with PAH (mPAP>20mmHg from cohort 1), sub-grouped by UCP2 genotype: WT (n=8), UCP2 HET SNP (n=6) and UCP2 HOM SNP (n=4). Scatter dot plots represent individual values. Quantification values (right) are expressed as mean ± SEM. E. UCP2 genotyping and echocardiography show the UCP2 genotype (blood) and Cardiac index in patients with PAH (mPAP>20mmHg from cohort 2). Patients were genotyped for UCP2 and categorized into WT (n=10), UCP2 HET SNP (n=6) and UCP2 HOM SNP (n=5). Scatter dot plots represent individual values. Quantification values ( right ) are expressed as mean ± SEM. *P < 0.05; **P < 0.01. F. UCP2 genotyping and echocardiography show the UCP2 genotype and Cardiac Index in patients with PAH (mPAP>20mmHg from cohort 2). Patients were genotyped for UCP2 and categorized into WT (n=10), UCP2 HET SNP (n=6) and UCP2 HOM SNP (n=5). Scatter dot plots represent individual values. Quantification values ( right ) are expressed as mean ± SEM. G. UCP2 genotyping and echocardiography show the UCP2 genotype and TAPSE in patients with PAH (mPAP>20mmHg from cohort 1and 2). Patients were genotyped for UCP2 and categorized into WT (n=16), UCP2 HET SNP (n=11) and UCP2 HOM SNP (n=9). Scatter dot plots represent individual values. Quantification values (right) are expressed as mean ± SEM *P < 0.05; **P < 0.01; ***P < 0.001. H. UCP2 genotyping and echocardiography show the UCP2 genotype and Cardiac index in patients with PAH (mPAP>20mmHg from cohort 1and 2). Patients were genotyped for UCP2 and categorized into WT (n=18), UCP2 HET SNP (n=12) and UCP2 HOM SNP (n=9). Scatter dot plots represent individual values. Quantification values (right) are expressed as mean ± SEM. *P < 0.05. I. UCP2 genotyping shows the ratio of patients with WT, UCP2-HET SNP and UCP2-HOM SNP in cRV vs dRV in patients with PAH (Cohort 1+2). J. UCP2 genotype and TAPSE in patients with sPHT (mPAP>20 mmHg from Cohort 3). Patients were genotyped for UCP2 and categorized into WT (n=6), UCP2 HET SNP (n=8) and UCP2 HOM SNP (n=3). Scatter dot plots represent individual values. Quantification values ( right ) are expressed as mean ± SEM. K. UCP2 genotype and TAPSE in patients with with sPHT (mPAP>20 mmHg from Cohort 3). Patients were genotyped for UCP2 and categorized into WT (n=8), UCP2 HET SNP (n=9) and UCP2 HOM SNP (n=4). Scatter dot plots represent individual values. Quantification values ( right ) are expressed as mean ± SEM. L. Spearman correlation test shows lack of orrelation between UCP2 RNA level and TAPSE in patients with sPHT (mPAP>20 mmHg from Cohort 3). Scatter dot plots show individual values. These comparisons were made using Kruskal–Wallis tests followed by pairwise Mann–Whitney U tests. SNP, Single Nucleotide Polymorphism; UCP2 , uncoupling protein 2; VDAC , Voltage-Dependent Anion Channels; WT, wild type; UCP2 HET, heterozygous UCP2 SNP; UCP2 HOM, homozygous UCP2 SNP; TAPSE, Tricuspid Annular Plane Systolic Excursion; mPAP: mean PA pressure; PAH, pulmonary arterial hypertension; s PHT : Secondary pulmonary hypertension; nRV, normal right ventricle; cRV, compensated right ventricle; dRV , decompensated right ventricle; cFB, cardiac fibroblasts; cMFB, cardiac myofibroblasts.

Techniques: Western Blot, MANN-WHITNEY

Journal: bioRxiv

Article Title: Estrogens protect bone mass by inhibiting NAD + metabolism in osteoclasts

doi: 10.1101/2025.07.11.664289

Figure Lengend Snippet: (a) Schematic illustration of NAD + biosynthetic pathways. In the de novo pathway the essential amino acid tryptophan (TRP) from the diet is utilized to produce NAD + via several reactions (dashed arrow). The salvage pathway recycles NAM, which is generated as a by-product of NAD + -dependent enzymes such as SIRT3; PA=Phthalic Acid. (b-h) BMMs were isolated from young female C57BL/6 mice and cultured with M-CSF and RANKL, in the presence or absence of FK866 or PA for 24 h. (b) NAD + levels in whole cells. (c) Representative images and quantification of TRAP-positive osteoclasts after 5 days. (d) NAD + levels in mitochondrial-enriched fractions. (e) SIRT3 activity. (f) Oxygen Consumption Rate. (g) ATP levels. (h) Apoptosis by caspase-3 activity. (i-j) Human BMMs were cultured with M-CSF and RANKL in the presence or absence of FK866, as above. (i) NAD + levels after 24 h and (j) representative images and quantification of TRAP-positive osteoclasts after 7 days. (k) mRNA levels of the indicated enzymes in BMMs cultured with M-CSF and RANKL the presence or absence of E 2 for the indicated timepoints. P values in grey indicate comparisons with time 0. P values in red indicate comparison of RANKL vs RANKL+E 2 within the same time point. (l) BMMs were isolated from young female C57BL/6 mice and cultured with M-CSF and RANKL the presence or absence of E 2 for 24 hours, representative Western blot images and quantification of the indicated proteins in mitochondrial enriched fractions; β-actin in cytosol and VDAC in mitochondria indicate efficacy of cellular compartment isolation. (m) mRNA levels in BMMs transduced with Nmnat1 and Nmnat3 sh or control sh lentivirus particles. (n) NAD + levels after 24 h and (o) representative images and quantification of TRAP-positive osteoclasts after 5 days. Scale bar 500 µm. Line and error bars represent mean ± SD. P values using two-tailed Student’s t-test, 1-way ANOVA followed by Dunnett’s multiple comparisons test, or 2-way ANOVA followed by Šídák’s multiple comparisons test.

Article Snippet: The following primary monoclonal antibodies were used: NAMPT (Abcam, ab236874, 1:1000); NMNAT3 (Santa Cruz, sc-390433, 1:100); FLAG (Cell Signaling; 2368; 1:1000); VDAC (Cell Signaling; D73D12; 1:1000) and β-actin (Santa Cruz; sc-47778; 1:5000).

Techniques: Generated, Isolation, Cell Culture, Activity Assay, Comparison, Western Blot, Transduction, Control, Two Tailed Test

Journal: bioRxiv

Article Title: Estrogens protect bone mass by inhibiting NAD + metabolism in osteoclasts

doi: 10.1101/2025.07.11.664289

Figure Lengend Snippet: (a) Chemical reaction catalyzed by Lb NOX. (b) Western blot for FLAG, β-actin, and VDAC from BMMs cultured with M-CSF expressing the FLAG-tagged cytosolic and mitochondrial (mito) Lb NOX. β-actin in cytosol and VDAC in mitochondria indicate efficacy of cellular compartment isolation. (c) NAD + /NADH redox ratio and (d) ATP levels in BMMs expressing Lb NOX or mito Lb NOX. (e-g) BMMs expressing Lb NOX or mito Lb NOX cultured with M-CSF and RANKL in the presence or absence of E 2 (10 -8 M) for 24 hours. (e) NAD + /NADH redox ratio. (f) SIRT3 activity. (g) Representative images and quantification of TRAP-positive osteoclasts after 5 days. Scale bar 500 µm. Line and error bars represent mean ± SD. P values by 1-way ANOVA followed by Dunnett’s multiple comparisons test or 2-way ANOVA followed by Šídák’s multiple comparisons test.

Techniques: Western Blot, Cell Culture, Expressing, Isolation, Activity Assay

Review

Structured Review

Images

1) Product Images from "Estrogens protect bone mass by inhibiting NAD + metabolism in osteoclasts"

Similar Products

Image Search Results