Journal: bioRxiv

Article Title: The Small Tumor Antigen of Merkel Cell Polyomavirus Accomplishes Cellular Transformation by Uniquely Localizing to the Nucleus Despite the Absence of a Known Nuclear Localization Signal

doi: 10.1101/2023.11.28.569067

Figure Lengend Snippet: Human foreskin fibroblasts (HFFs) were transduced with MCPyV ST or GFP, followed by TurboID Mass Spectrometry analysis. A total of 121 proteins were enriched in ST expressing HFFs defined as a False Discovery Rate (FDR) <0.01 and Fold Change (FC) > 2. These 121 proteins were colored based on their subcellular localization, with teal representing nuclear proteins, pink representing cytoplasmic proteins, and green representing proteins with both nuclear and cytoplasmic localization (A). A Venn diagram was also included to visualize the number of MCPyV ST potential interactors that are nuclear, cytoplasmic, or both nuclear and cytoplasmic (B). The amino acid sequence of MCPyV ST contains no known canonical NLS (C). Online amino acid and protein subcellular localization predictor tools were utilized to predict the localization of MCPyV ST (D). The location of the MCPyV LT-t NLS is found within the MCPyV LT-t unique region (orange), but no known NLS is found within the common-T region (red) or ST unique region (blue) (E).

Article Snippet: Adenovirus-transformed human embryonic kidney cells (HEK293A), human foreskin fibroblasts (HFF), 293TNs, and Rat-2 cells were cultured in 1x DMEM (ThermoFisher Scientific) supplemented with 10% fetal bovine serum (FBS) (Neuromics), GlutaMax (ThermoFisher Scientific), Non-Essential Amino Acids (NEAA) (ThermoFisher Scientific), and 100 units/mL penicillin-streptomycin (ThermoFisher Scientific).

Techniques: Transduction, Mass Spectrometry, Expressing, Sequencing

Journal: bioRxiv

Article Title: Pyrimidine salvage in Toxoplasma gondii as a target for new treatment

doi: 10.1101/2023.11.01.565095

Figure Lengend Snippet:

Article Snippet: The in vitro inhibitory effects of antimicrobial nucleoside agents on T. gondii growth were determined in HFF cell cultures using 96-well black plates (Optical-Bottom Plates with Polymer Base, ThermoFisher Scientific).

Techniques: In Vitro

Journal: Pathogens

Article Title: Functional Characterization of Eight Zinc Finger Motif-Containing Proteins in Toxoplasma gondii Type I RH Strain Using the CRISPR-Cas9 System

doi: 10.3390/pathogens12101232

Figure Lengend Snippet: Invasion, intracellular replication, egress, and parasite virulence assays of RHΔ zfp strains and WT strains. ( A ) Compared with the WT strain, the invasion efficiency of eight RHΔ zfp strains showed no significant reduction. ( B ) HFF monolayers were infected with RHΔ zfp strains and the WT strain for 24 h. Parasitophorous vacuoles (PVs) containing 1, 2, 4, 8, and 16 tachyzoites were counted from 200 randomly selected PVs. Only the replication process of RHΔ 273150 showed significant inhibition compared to the WT strain. ( C ) No differences were detected in the egress efficiency between the eight RHΔ zfp strains and the WT strain. ***, p < 0.001, n.s., not significant.

Article Snippet: For the plaque assay, HFF monolayers in a 12-well cell plate (Thermo Fisher Scientific) were infected with freshly egressed RHΔ zfp strains and WT strain (approximately 500 tachyzoites per well).

Techniques: Infection, Inhibition