Journal: International Journal of Molecular Sciences

Article Title: Activation of ERK and p38 Reduces AZD8055-Mediated Inhibition of Protein Synthesis in Hepatocellular Carcinoma HepG2 Cell Line

doi: 10.3390/ijms222111824

Figure Lengend Snippet: Screening anticancer drugs for identification of translation inhibitors. ( a , b ) The level of translation in mouse embryonic fibroblasts (MEFs) was measured using puromycin incorporation assay. Polypeptide chains in wild-type MEFs treated with 10 μM ( a ) or the indicated concentrations ( b ) of Selleck Anti-cancer Compound Library for 24 h were labeled with puromycin for 10 min. The number of cells (left column) and Alexa Fluor 488 intensities of each area (right column) were assessed using Operetta High-Content analysis system. Operetta images were analyzed using Harmony software. Red dots in the red dotted circle in B indicate MEFs treated with various concentrations of AZD8055. To recognize the shape of cells, cells were stained with Hoechst 33342 prior to the measurement of cytoplasmic Alexa Fluor 488 intensities. The number of compounds include DMSO- and Tg-treated samples. ( c – e ) Wild-type (S/S) or eIF2α mutant (A/A) MEFs were treated with the indicated concentrations of AZD8055 for 24 h; two representative confocal images are shown ( c ). The intensity of Alexa Fluor 488 was visualized ( c ) and MEFs treated with various concentrations of AZD8055 were magnified ( c ). Scale bar = 200 μM. The obtained images were measured using Harmony Software ( d ). Immunoblot analyses were performed to determine the relative amount of puromycin and phosphorylated form of eIF2α using specific antibodies for puromycin, eIF2α, phosphorylated form of eIF2α (p-eIF2α), and GAPDH ( e ). Treatment with thapsigargin (Tg), an ER stress inducer, was used as a positive control. Fold changes in d were calculated by comparing DMSO control group of each cell line with AZD8055 treatment groups. Statistically significant differences were determined by Student’s t -test and were indicated as *** p < 0.001.

Article Snippet: The cells were treated with dimethyl sulfoxide (DMSO; Biosesang, Korea) or Selleck Anti-Cancer Compound Library for 24 h. Cells treated with 1 µM Tg for 30 min served as positive control.

Techniques: Labeling, High Content Screening, Software, Staining, Mutagenesis, Western Blot, Positive Control

Journal: Emerging microbes & infections

Article Title: Identification of cepharanthine as an effective inhibitor of African swine fever virus replication.

doi: 10.1080/22221751.2024.2429624

Figure Lengend Snippet: Figure 1. Screening of antiviral compounds against ASFV in the Selleck natural compounds library. (A) Depiction of the screening process of anti-ASFV compounds. A total of 803 compounds were screened in ASFV infected PAMs (1 MOI) using a cell-based Elisa assay. Two replicates were set for each compound at a concentration of 10 μM, and those with high cytotoxicity were excluded in the second round of screening. (B) The screening results of anti-ASFV compounds. Each dot represents a compound, ranging by infection ratio. (C) The 50% cytotoxicity concentrations (CC50) of candidate anti-ASFV compounds. PAMs (2 × 105 cells/well) were seeded in a 96 well plate overnight, treated with the indicated concentration of DMSO or compounds for 24 h, and then treated with CCK-8 agent for an additional 2 h. The OD450 value was measured for cell viability detection. (D) The Half Maximal Inhibitory Concentration (IC50) of candidate anti-ASFV compounds. PAMs were treated with the indicated concentration of DMSO or com pounds for 2 h, followed by a cell-based ELISA assay. Data were calculated through nonlinear regression analysis.

Article Snippet: Screening of antiviral compounds against ASFV in the Selleck natural compounds library. (A) Depiction of the screening process of anti-ASFV compounds.

Techniques: Infection, In-Cell ELISA, Concentration Assay, CCK-8 Assay