Journal: Journal of Radiation Research

Article Title: Mitochondrial-targeted SS-31 peptide attenuates radiation-induced cardiomyocyte senescence

doi: 10.1093/jrr/rrag048

Figure Lengend Snippet: SS-31 reduces radiation-induced mitochondrial-related apoptosis proteins in H9C2 cells and hiPSC-CMs . (A, C) Representative western blot images showing the expression of mitochondrial apoptosis-related proteins Bax and Bcl-2 in H9C2 cells (A) and hiPSC-CMs (C). (B, D) Quantification of corresponding protein expression levels, n = 3–10. Experimental groups include control, SS-31 treatment, 10 Gy radiation, and 10 Gy radiation plus SS-31 treatment, as indicated in the figure. (E) Representative western blot images of cytochrome c (Cyt c) distribution. Protein levels were analyzed in isolated mitochondrial and cytosolic fractions. (F) Quantitative analysis of Cyt c expression. Cyt c levels were normalized to VDAC1 for the mitochondrial fraction and β-actin for the cytosolic fraction. n = 3–4. All data are presented as mean ± SEM. * P < 0.05 vs control; ** P < 0.01 vs control; # P < 0.05 vs irradiation; ## P < 0.01 vs irradiation.

Article Snippet: Membranes were blocked with 1% BSA or 1% non-fat milk in tris-buffered saline with Tween 20 (TBST) for 1 hour at room temperature, followed by overnight incubation at 4°C with the following primary antibodies: TNFα (MILLIPORE, AB1837P, 1:2000), IL-6 (Sigma, SAB5700632, 1:2000), p16 (Invitrogen, PA5–20379, 1:2000), p21 (Invitrogen, 14–6715-81, 1:2000), Bax (Invitrogen MA5–14003, 1:1000), Bcl-2 (Abcam, ab196495, 1:2000), Lamin B1 (CST, 13435, 1:2000), Caspase 7 (CST, 9492S, 1:2000), Caspase 8 (CST, 4790S, 1:2000), Caspase 9 (CST, 9508S, 1:1000), Cytochrome c (Medchemexpress, HY- P80102 , 1:3000), VDAC1 (Medchemexpress, HY- P80369 , 1:3000), OxPhos Rodent WB Antibody Cocktail (Invitrogen, 45–8099, 1:2000), or β-actin (Bio-techne, MAB8929, 1:5000).

Techniques: Western Blot, Expressing, Control, Isolation, Irradiation

Journal: Alzheimer's & Dementia

Article Title: Endothelial NAD + depletion drives vascular senescence and neuroinflammation via mtDNA‐cGAS/STING‐CD38 signaling in Alzheimer's disease

doi: 10.1002/alz.71423

Figure Lengend Snippet: NAD + restoration prevents VDAC1 oligomerization‐mediated mtDNA leakage and cGAS/STING pathway activation in AD. (A, B) Enzymatic activities of isocitrate dehydrogenase (ICDH) (A; n = 6 per group) and α‐ketoglutarate dehydrogenase (α‐KGDH) (B; n ≥5 per group) in mitochondrial‐enriched fractions from cerebral vessel‐enriched fractions of APPwt, APPwt + NR, APPtg, and APPtg + NR mice. (C) Citrate accumulation in mitochondrial‐enriched fractions from cerebral vessel‐enriched fractions of APP/PS1 mice, which was significantly increased compared to APPwt controls and normalized following NAD + restoration ( n ≥5 per group). (D) Immunoblot analysis showing enhanced VDAC1 oligomerization in cerebral vessel‐enriched fractions from APP/PS1 mice, which was markedly suppressed by NR treatment. (E) Quantification of cytosolic mtDNA fragments ( D‐loop , Non‐Numt , and Cox1 ) in Aβ‐treated bEnd.3 endothelial cells with or without VBIT‐4 (VDAC1 oligomerization inhibitor) treatment ( n = 4 per group). (F) Representative immunoblot showing VDAC1 oligomerization in Aβ‐treated bEnd.3 cells, which was suppressed by NR or VBIT‐4 treatment. (G) Immunoblot and densitometric quantification showing decreased expression of cGAS, STING, and phosphorylated TBK1 and IRF3 following VBIT‐4 treatment in Aβ‐treated bEnd.3 cells ( n = 4 per group). Data are presented as mean ± SEM. Statistical analyses were performed using one‐way ANOVA followed by Tukey's multiple comparisons test. P ‐values are indicated in the figure.

Article Snippet: To specifically inhibit VDAC1 oligomerization, bEnd.3 cells were pretreated with VBIT‐4 (10 μM; MedChemExpress, #HY‐101966) for 24 h in serum‐reduced medium before being subjected to Aβ challenge.

Techniques: Activation Assay, Western Blot, Expressing