vdac  (Cell Signaling Technology Inc)

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

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: 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.

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: Western Blot, Control, Injection

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. 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.

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: Western Blot, MANN-WHITNEY