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: Biological & pharmaceutical bulletin

Article Title: Changes in PtdIns(4,5)P2 induced by etoposide treatment modulates small intestinal P-glycoprotein via radixin.

doi: 10.1248/bpb.b13-00953

Figure Lengend Snippet: Fig. 1. Effect of Rosuvastatin or Fasudil on the Increased PtdIns(4,5)P2 Expressions in the Ileal Membrane after Repeated Oral Treatment with ETP

Article Snippet: Drug Administration The drugs used in this study were etoposide phosphate (ETP; Sequoia Research Products, Pangbourne, U.K.), rosuvastatin calcium (rosuvastatin; Toronto Research Chemicals Inc., North York, ON, Canada) and fasudil hydrochloride (fasudil; Tocris Bioscience, Bristol, U.K.).

Techniques: Membrane

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: Vehicle-dependent Effects of Sphingosine 1-phosphate on Plasminogen Activator Inhibitor-1 Expression

doi: 10.5551/jat.37663

Figure Lengend Snippet: S1P bound to albumin induced PAI-1 expression by activating S1PR2 pathway in 3T3L1 adipocytes. (A) 3T3L1 adipocytes were pre-incubated for 18 hours in FBS-free DMEM and the medium was exchanged for FBS-free DMEM containing 20 µM of VPC23019, JTE013, or DMSO for 30 minutes. Then, the cells were challenged with a medium containing 10 µM of sphingosine 1-phosphate (S1P) with the corresponding inhibitors for 4 hours. The mRNA level of plasminogen activator inhibitor 1 (PAI-1) was analyzed using real-time PCR. 18S was utilized as an internal control ( n = 4–6/group). † p < 0.01 vs. DMSO alone, YC1 alone, S1P + VPC, and JTE alone, and p < 0.05 vs. S1P + JTE. ‡ p < 0.01 vs. other groups. (B, C). 3T3L1 adipocytes were pre-incubated for 18 hours in FBS-free DMEM, and the medium was exchanged for FBS-free DMEM with 10 µM of S1P bound to apoM-rich vehicle (ApoM) or control vehicle (Null) for 4 hours. Then, the modulation of the phosphorylation of moesin in 3T3L1 cells was examined using western blotting ( n = 6/group). ** p < 0.01, *** p < 0.001.

Article Snippet: VPC23019 (857360P; Avanti Polar Lipids, Alabaster, AL), JTE013 (10009458; Cayman Chemical, Ann Arbor, MI), Y27632 (257-00511; WAKO Pure Chemical Industries, Osaka, Japan), wortmannin, YC-1 (W1628, Y102; Sigma-Aldrich Co), and SIS3 and BAY11-7082 (sc-222318, sc-200615; Santa Cruz Biotechnology, Inc. TX) were dissolved in DMSO.

Techniques: Expressing, Incubation, Real-time Polymerase Chain Reaction, Control, Phospho-proteomics, Western Blot