In vitro evaluation of the efficacy of azithromycin against the larvae of Echinococcus multilocularis
Description
Western blot
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Steps to reproduce
Proteins from EmPSCs were extracted using a Total Protein Extraction Kit (Solarbio, China), and protein concentrations were measured using a BCA protein assay (Thermo Fisher Scientific, USA). Subsequently, the protein samples were separated using 10% SDS-PAGE for 2 h, transferred to PVDF membranes, and blocked with 5% skim milk for 1 h. After washing, and then incubate with Anti-AKT (Cell Signaling Technology, 2920, 1:2000), Anti-Phospho-AKT(Cell Signaling Technology, 4058, 1:2000), Anti-Glucose Transporter GLUT1 antibody(Abcam, ab115730, 1:5000), anti-gamma H2AX (Abcam, ab81299, 1:7500), DHODH (Proteintech, 14877-1-AP, 1:8000), and GPX4 (Affinity, DF6701, 1:2000) antibodies at 4°C overnight. After washing, the membranes were incubated with goat anti-rabbit IgG/HRP (Bioss, bs-80295G-HRP, 1:5,000) or goat anti-mouse IgG/HRP (Bioss, bs-0296G-HRP, 1:5,000) secondary antibodies at room temperature for 1 h. Protein expression was quantified by measuring the grayscale value of each band using ImageJ software, with normalization to β-actin. Relative protein expression for each biological replicate was then calculated as the fold change relative to the control group. Representative blots from at least three independent experiments are shown in the figures. The data are presented as mean±SD obtained from three independent experiments.