Journal: bioRxiv

Article Title: The Legionella pneumophila type IVb secretion system effector BinA subverts amino acid transport to sensitize TORC1 signaling in macrophages

doi: 10.1101/2025.02.20.639245

Figure Lengend Snippet: (A) Schematic depiction of the LAT1-dependent uptake assay for import of the amino acid analog π-Boronophenylalanine (L-BPA). Cell were treated with L-BPA for 5min (1) after which cells were extensively washed, followed by the addition of a specific membrane permeable probe for additional 5 min (2), which emits fluorescent signal when bound to L-BPA(3). (B) Graph shows L- BPA uptake by Myd88 KO iBMDMs infected with the indicated Lp strains at MOI=20 or left uninfected (UN). In some control conditions, the LAT1 inhibitor 2-Aminobicyclo [2.2.1] heptane-2-carboxylic acid (BCH) at 1mM was added 10 min prior to and for the duration of the L-BPA uptake assay. BPA uptake lasted 5min and was measured at 4 hpi, after which fluorescence output was measured, the cells were lysed and total protein content was measured. Mean fluorescence data was normalized to the total protein content. Averages ± SEM of data from three biological repeats are shown for each condition. Statistical analysis was completed via one-way ANOVA Kruskal-Wallis test with Dunn’s multiple comparison and the respective p-values among the conditions compared are indicated.

Article Snippet: BPA uptake assay (Dojindo Labratories, cat# UP04) was used to measure BPA transport in Lp-infected macrophages as per the suggested manufacturer’s protocol.

Techniques: Membrane, Infection, Control, Fluorescence, Comparison

Journal: Drug Metabolism and Disposition

Article Title: The Presence of a Transporter-Induced Protein Binding Shift: A New Explanation for Protein-Facilitated Uptake and Improvement for In Vitro-In Vivo Extrapolation

doi: 10.1124/dmd.118.085779

Figure Lengend Snippet: Uptake curves for pravastatin (A), rosuvastatin (B), atorvastatin (C), and pitavastatin (D). Total uptake is depicted in circles, passive diffusion is depicted in squares, and active uptake is depicted in triangles. The error bars represent the S.D. of the triplicate.

Article Snippet: Atorvastatin was purchased from TCI America (Portland, OR), pitavastatin from ApexBio (Houston, TX), rosuvastatin from Toronto Research Chemicals (North York, ON, Canada), and [3H(G)] pravastatin sodium salt (specific activity, 5 Ci/mmol) from American Radiolabeled Compounds (St. Louis, MO).

Techniques: Diffusion-based Assay

Journal: Drug Metabolism and Disposition

Article Title: The Presence of a Transporter-Induced Protein Binding Shift: A New Explanation for Protein-Facilitated Uptake and Improvement for In Vitro-In Vivo Extrapolation

doi: 10.1124/dmd.118.085779

Figure Lengend Snippet: K m,u , V max , P dif,u , and CL int values generated for each compound in buffer and plasma incubations

Article Snippet: Atorvastatin was purchased from TCI America (Portland, OR), pitavastatin from ApexBio (Houston, TX), rosuvastatin from Toronto Research Chemicals (North York, ON, Canada), and [3H(G)] pravastatin sodium salt (specific activity, 5 Ci/mmol) from American Radiolabeled Compounds (St. Louis, MO).

Techniques: Generated, Clinical Proteomics, Incubation