Journal: The Journal of general virology

Article Title: Influenza virus entry via the GM3 ganglioside-mediated platelet-derived growth factor receptor β signalling pathway.

doi: 10.1099/jgv.0.001235

Figure Lengend Snippet: Fig. 4. Virus entry and inhibition by Ki8751 in CHO-wt, CHO-K1, and transfectant cells carrying FGFR4 or VEGFR2. (a) Nuclear import of vRNP in CHO cells incubated with influenza (A/PR/8/34) virus over 1 h at 35 C. Quantitative confocal microscopy was carried out after dual nuclear staining for viral NP and DAPI. In three separate experiments, three images (each containing ~120 cells) were taken from each condition to quantify the green fluorescent signal in the nucleus. (b) Virus binding efficiency relative to CHO-wt cells, as determined by RT-qPCR quantification of cell-bound virus after 60 min of incubation at 4 C. (c) Ki8751 and two other RTK inhibitors, crenolanib and imatinib (all at 60 µM), inhibit virus nuclear entry in CHO-wt and CHO-K1 cells. Confocal microscopy after NP-immunos- taining at 1 h p.i. Green: viral NP; blue: nuclear DAPI staining. (d) vRNA synthesis at 10 h p.i., assessed by two-step RT-qPCR. The y-axis shows the fold increase in vRNA, relative to untreated CHO-wt cells. Panels a, b and d: mean±SEM (N=3). Statistical significance by two- sided Student’s t-test: *P<0.05; ***P<0.005; NS=not significant (P>0.05).

Article Snippet: The library of 276 protein kinase inhibitors was purchased from Selleck Chemicals (catalogue no. L1200).

Techniques: Virus, Inhibition, Transfection, Incubation, Confocal Microscopy, Staining, Binding Assay, Quantitative RT-PCR

Journal: Frontiers in Pharmacology

Article Title: Nephrotoxicity of Calcineurin Inhibitors in Kidney Epithelial Cells is Independent of NFAT Signaling

doi: 10.3389/fphar.2021.789080

Figure Lengend Snippet: Effects of CsA on kidney epithelial cells are mimicked by protein kinase inhibitors. (A-B) Protein kinase library screen revealed that inhibition of PI3K/Akt, IKK, and p38 MAPK induces Fn14 (MCT). (C) Titration of two p38 MAPK inhibitors (SB203580 and SB202190) revealed the similar potential to induce Fn14 (MCT). (D) Protein levels of Fn14 are increased upon CsA and SB203580 treatment but not when NFAT is inhibited (MCT, Western Blot). (E) Increased expression of Fn14 is detected when cells are treated simultaneously with the p38 MAPK inhibitor and CsA compared to single treatment with inhibitors (MCT cells). (F) Constitutive active p38 MAPK shows reduced Fn14 expression on cells when treated with either SB203580 or CsA (HEK293T). (G) Nephrotoxic genes are upregulated with CsA and with SB203580 (MCT, RT-PCR). If not indicated differently, results were acquired by flow cytometry.

Article Snippet: The next day, cells were treated with various kinase inhibitors (Enzo, BML-2832) at 10 μM for 48 h and then analyzed by flow cytometry (described below).

Techniques: Inhibition, Titration, Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: Nephrotoxicity of Calcineurin Inhibitors in Kidney Epithelial Cells is Independent of NFAT Signaling

doi: 10.3389/fphar.2021.789080

Figure Lengend Snippet: In vivo experiments show the development of nephrotoxicity upon CsA and p38-kinase inhibitor. (A) Nephrotoxic gene array reveals evident upregulation of genes involved in nephrotoxicity upon CsA treatment. Also, both p38 inhibitors revealed upregulation of nephrotoxic genes (mouse kidney cells, RNAseq). (B) Higher CD45 +- infiltrated cells in the kidneys of CsA, SB203580, and SB202190 were detected. (C) Monocytic and lymphocytic cell compartments in the blood show a slight increase in cell number in animals treated with CsA and p38 MAPK inhibitors. B and C were measured by flow cytometry.

Article Snippet: The next day, cells were treated with various kinase inhibitors (Enzo, BML-2832) at 10 μM for 48 h and then analyzed by flow cytometry (described below).

Techniques: In Vivo, Flow Cytometry