Journal: The Journal of Biological Chemistry

Article Title: Converging Evidence of Mitochondrial Dysfunction in a Yeast Model of Homocysteine Metabolism Imbalance *

doi: 10.1074/jbc.M111.228072

Figure Lengend Snippet: ER stress response in str4Δ in the presence of homocysteine and its precursors. A, unfolded protein response in str4Δ cells. Cells were transformed with a plasmid (pJC005), a 2-μm plasmid carrying the lacZ gene under the control of the UPRE from the KAR2 promoter. Transformed cells were grown in respective conditions; cells were harvested at 16 h, and β-galactosidase activity was measured using 2-nitrophenyl-β-d-galactopyranoside as a substrate and detected as an increase in absorbance at 420 nm. B, expression of KAR2 gene determined by Northern blotting. The cells were grown in the presence and absence of homocysteine and its precursors for 16 h, and RNA was isolated from these cells. 20 μg of RNA was loaded per lane for Northern blot analysis. The lower panels in each blot represent gel loading of total RNA. Image was acquired using PhosphorImager and quantified by ImageQuant software. SAM, AdoMet; SAH, AdoHcy.

Article Snippet: To compare lacZ expression in WT yeast versus str4 mutant yeast under quiescent or ER stress conditions, the β-galactosidase activity was measured from lysates using β-galactosidase assay kit (Promega).

Techniques: Transformation Assay, Plasmid Preparation, Activity Assay, Expressing, Northern Blot, Isolation, Software

Journal: bioRxiv

Article Title: Radiation-induced non-neoplastic cells senescence promoting the proliferation and invasiveness of GBM through SASP/JAK2-STAT3 Pathway

doi: 10.1101/2024.02.21.581353

Figure Lengend Snippet: (A) β-galactosidase staining on NHA cells 7 days after radiation and mock-radiation. (B) mRNA expression levels of P16 and P21 genes in U251 and A172. (C , D) β-galactosidase staining of mouse brain tissue at different time points after radiation and trend diagram of senescent cell number. (E) β-galactosidase staining of human tumor specimens. Cases 1,2 and 3 were primary GBM; Cases 4,5, and 6 were recurrent GBM and received radiotherapy.

Article Snippet: The experiment was conducted following the protocol of β-galactosidase staining kit (Solarbio, G1580).

Techniques: Staining, Expressing

Journal: International Journal of Nanomedicine

Article Title: Combined Placental Mesenchymal Stem Cells with Guided Nanoparticles Effective Against Diabetic Nephropathy in Mouse Model

doi: 10.2147/IJN.S446733

Figure Lengend Snippet: Internalization of Nanoparticles (NPs) into Placental Mesenchymal Stem Cells (PL-MSCs). ( A ) MSCs stained with Prussian Blue to highlight iron (Fe) content. Scale bar = 50 µm. ( B ) Quantification of Fe concentration using Inductively Coupled Plasma-Optical Emission Spectroscopy (ICP-OES) (n=3). ( C ) Assessment of NP-induced cytotoxicity in MSCs labeled with NPs at varying concentrations for 24 h (n=4). ( D ) Measurement of β-galactosidase levels in NP-labeled MSCs using ELISA (n=4). ( E ) Proliferation rates of cells labeled with NPs at different concentrations (n=5). ( F and G ) Transmission Electron Microscopy (TEM) images showing NPs internalized into an MSC at a concentration of 50 µg/mL. The red arrow indicates NP. Data are presented as mean ± S.E.M. P -values < 0.05 were considered statistically significant, denoted as * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: The supernatants were collected for β-galactosidase analysis using a human β-galactosidase ELISA kit (Molbio, Shanghai, China), in accordance with the manufacturer’s protocol.

Techniques: Staining, Concentration Assay, Clinical Proteomics, Spectroscopy, Labeling, Enzyme-linked Immunosorbent Assay, Transmission Assay, Electron Microscopy