Journal: Genetics and molecular research : GMR

Article Title: Increased survivin expression and its association with clinical parameters of congenital choledochal cysts.

doi: 10.4238/gmr.15028032

Figure Lengend Snippet: Figure 1. Immunohistochemical staining results for the protein expression of survivin. The cytoplasm showed high survivin expression, which presented as granular form, with yellow or dark brown staining, while the nuclei were hardly stained (A. cytoplasm, 200X; B. nuclei, 400X).

Article Snippet: The primary mouse anti-human survivin monoclonal antibody was purchased from Wuhan Boster Biological Engineering Co., Ltd. (China).

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Theranostics

Article Title: Progerin accumulation in nucleus pulposus cells impairs mitochondrial function and induces intervertebral disc degeneration and therapeutic effects of sulforaphane.

doi: 10.7150/thno.30658

Figure Lengend Snippet: Figure 3. Progerin induces aging-related defects and mitochondrial dysfunction in NP cells. (A) Representative IF images and quantification of LAP2, γH2AX, H3K27me3, and lamin B1 expression in the vector con and progerin groups; n = 5; **P < 0.01. (B) TUNEL staining of NP cells in the vector con and progerin groups. Nuclei were stained with DAPI. The number of TUNEL-positive cells was significantly greater in the progerin group than in the vector con group; n = 5; **P < 0.01. (C) Representative flow cytometry dot plots of apoptosis after Annexin V-FITC/PI dual staining. The relative number of apoptotic cells was greater in the progerin group compared to the vector con group; n = 5; **P < 0.01. (D) Representative images and quantification of DCFDA-based ROS levels in the vector con and progerin groups; n = 5; **P < 0.01. (E) JC-1 staining. The red: green fluorescence ratio reflects changes in the mitochondrial membrane potential of NP cells in the vector con and progerin groups; n = 5; **P < 0.01. (F) ATP production in the vector con and progerin groups; n = 5; *P < 0.05. (G) Relative activities of mitochondrial complex enzymes in NP cells in the vector con and progerin groups; n = 4; *P < 0.05, **P < 0.01. Data represent mean ± SEM. LAP2, lamina-associated polypeptide2; γH2AX, serine-139 phosphorylated H2AX; H3K27me3, heterochromatin-associated tri-methylated lysine 27 on histone 3; SA-β-gal, senescence-associated β-galactosidase; DCFDA, 2',7'-dichloro-dihydrofluorescein-diacetate; JC-1, 5,5-,6,6-tetrachloro-1,1-,3,3-tetraethylbenzimidazole-carbocyanine iodide.

Article Snippet: For immunofluorescence and immunohistochemical analyses, the following antibodies were used: Anti-aggrecan (rabbit polyclonal; Proteintech, #13880-1-AP), P16INK4a mouse monoclonal (Abcam, #ab54210), LAP2 rabbit polyclonal (Proteintech, #14651-1-AP), γH2AX (Proteintech, #10856-1-AP), H3K27me3 rabbit monoclonal (CST, #9733), lamin B1 rabbit polyclonal (Proteintech, #12987-1-AP), progerin mouse monoclonal (Santa Cruz, #sc-81611), and collagen II rabbit polyclonal (Abcam, #ab34712).

Techniques: Expressing, Plasmid Preparation, TUNEL Assay, Staining, Cytometry, Membrane, Methylation

Journal: Theranostics

Article Title: Progerin accumulation in nucleus pulposus cells impairs mitochondrial function and induces intervertebral disc degeneration and therapeutic effects of sulforaphane.

doi: 10.7150/thno.30658

Figure Lengend Snippet: Figure 5. SFN ameliorates progerin-induced aging defects and mitochondrial dysfunction. (A) IF of LAP2, γH2AX, H3K27me3, and lamin B1 in the progerin+vehicle and progerin+SFN groups; n = 5; *P <0.05. (B) Representative images and quantification of ROS levels in the progerin+vehicle and progerin+SFN groups; n = 5; *P < 0.05. (C) JC-1 staining. The red: green fluorescence ratio reflects changes in the mitochondrial membrane potential of NP cells in the progerin+vehicle and progerin+SFN groups; n = 5; *P < 0.05. (D) ATP production in the progerin+vehicle and progerin+SFN groups; n = 5; *P < 0.05. (E) Western blotting analysis of OPA1, Drp1, Mfn1, and Mfn2 in NP cells from the vector+vehicle, progerin+vehicle, and progerin+SFN groups; n = 5; *P < 0.05, **P < 0.01. (F) Western blotting analysis of PGC1α, pAMPK, and AMPK in NP cells from the vector+vehicle, progerin+vehicle, and progerin+SFN groups; n = 5. β-actin was used as the control. *P < 0.05, **P < 0.01. Data represent mean ± SEM. SA-β-gal, senescence-associated β-galactosidase; DCFDA, 2',7'-dichloro-dihydrofluorescein-diacetate; JC-1, 5,5-,6,6-tetrachloro-1,1-,3,3-tetraethylbenzimidazole-carbocyanine iodide; OPA1, optic atrophy-1; Drp1, dynamin-related peptide1; Mfn1, mitofusin1; Mfn2, mitofusin2; PGC1α, peroxisome proliferator-activated receptor-γ coactivator1α; AMPK, AMP-activated protein kinase.

Article Snippet: For immunofluorescence and immunohistochemical analyses, the following antibodies were used: Anti-aggrecan (rabbit polyclonal; Proteintech, #13880-1-AP), P16INK4a mouse monoclonal (Abcam, #ab54210), LAP2 rabbit polyclonal (Proteintech, #14651-1-AP), γH2AX (Proteintech, #10856-1-AP), H3K27me3 rabbit monoclonal (CST, #9733), lamin B1 rabbit polyclonal (Proteintech, #12987-1-AP), progerin mouse monoclonal (Santa Cruz, #sc-81611), and collagen II rabbit polyclonal (Abcam, #ab34712).

Techniques: Staining, Membrane, Western Blot, Plasmid Preparation, Control

Journal: Frontiers in Cardiovascular Medicine

Article Title: Momordica. charantia -Derived Extracellular Vesicles-Like Nanovesicles Protect Cardiomyocytes Against Radiation Injury via Attenuating DNA Damage and Mitochondria Dysfunction

doi: 10.3389/fcvm.2022.864188

Figure Lengend Snippet: MCELNs ameliorated mitochondrial dysfunction induced cell injury after radiation. (A) H9C2 cells after 48 h of culture with indicated treatment were incubated with mitochondria ROS probe (red) for 15 min. Then nucleus were stained with DAPI (blue) for confocal imaging, scale bar: 20 μm. Flow analysis (B) and quantitation (C) of mitochondria ROS intensity in H9C2 cells after 48 h of culture with indicated treatment. Flow analysis (D) and quantitation (E) of mitochondria membrane potential in H9C2 cells after 48 h of culture with indicated treatment that detected by JC-1 probe. Western blot images (F) and quantitation (G) of AKT, p-AKT, ERK, and p-ERK in H9C2 cells after 48 h of culture with indicated treatment. IR (–/+): 0/16 Gy X-ray; MCELNs (–/+): 0/10 μg/mL. All data were represented as means ± SD ( n = 3 independent experiments). The statistical significance was evaluated by one-way ANOVA followed by the Turkey's multiple comparisons test among groups.

Article Snippet: The primary antibodies included TSG101 (#28283-1-AP, proteintech), CD63 (#25682-1-AP, proteintech), CD9 (#20597-1-AP, proteintech), PCNA (#A0264, ABclonal), Cyclin D1 (#2978, CST), Cyclin B1 (#55004-1-AP, proteintech), cleaved caspase3 (#9664, CST), cleaved PARP (#9548, CST), γ-H2A.x (#ab2893, Abcam), ATM (#NB100-309, NOVUS), p-ATM (#NB100-306, NOVUS), AKT (#60203-2-lg, proteintech), p-AKT (#4060, CST), ERK (#BF8004, affinity), p-ERK (#AF1015, affinity), β-actin (#66009-1-lg, proteintech), α-tubulin (#66031-1-lg, proteintech).

Techniques: Incubation, Staining, Imaging, Quantitation Assay, Membrane, Western Blot