Journal: Medical Sciences
Article Title: Combined Effects of Fludarabine and Interferon Alpha on Autophagy Regulation Define the Phase of Cell Survival and Promotes Responses in LLC-MK2 and K562 Cells
doi: 10.3390/medsci10010020
Figure Lengend Snippet: Western blot analysis of autophagy response and the antiviral response in cultured LLC-MK2 and K562 cells that were exposed for 60 min to IFN-α (10 U/mL) before treatment with Fdb (10 μM) for 24 h. The cells were evaluated for autophagy response mediation via p-mTOR and LC3B-I/LC3B-II activation, as well as antiviral response via p-STAT1 signaling molecules. The total proteins were extracted and transferred to nitrocellulose membranes. The membranes were immunostained with p-mTOR, LC3B-I/LC3B-II, and p-STAT1 antibodies and processed for reaction color development. β-Actin was used as an internal control. Protein electrophoresis films were digitized and the relative density level of p-mTOR (normalized with β-Actin), autophagosome marker LC3B-II (normalized with LC3B-I), and p-STAT1 (normalized with β-Actin) were measured by using ImageJ ® software. One-way ANOVA statistical analysis was performed (*, **, *** indicates a p -value of <0.05, 0.01, 0.001, respectively). The experiments were analyzed in triplicate and SEM is marked by an error bar.
Article Snippet: LLC-MK2, the rhesus monkey kidney epithelial cell line, and the K562 human leukemic cell line were purchased from ATCC (Manassas, VA, USA), propagated, and stocked at −80 °C.
Techniques: Western Blot, Cell Culture, Activation Assay, Control, Protein Electrophoresis, Marker, Software