Journal: Journal of Virology

Article Title: Newcastle disease virus activates diverse signaling pathways via Src to facilitate virus entry into host macrophages

doi: 10.1128/jvi.01915-23

Figure Lengend Snippet: PI3K/AKT downstream Rac1-PAK1 signaling axis is not activated and involved in NDV entry into HD11 cells. (A) GTPase activity of Rac1 was analyzed by using the Rac1 activation assay kit, followed by Western blotting using an anti-Rac1 antibody. The level of GTP-Rac1 was normalized to the total Rac1. (B) The levels of p-PAK1 (T423) and PAK1 were analyzed by Western blotting using corresponding antibodies. GAPDH was used as a control. (C–G) Treatment with the Rac1 and PAK1 inhibitor, NSC23766 and IPA-3, showed no effect on NDV-induced LIMK1/CFN phosphorylation and entry of NDV. (C) HD11 cells were pretreated with NSC23766, IPA-3, or DMSO. Then the cells were inoculated or mock inoculated with NDV. Western blotting was used to determine the levels of p-LIMK1 (T508), LIMK1, p-CFN (S3), and CFN at 30 mpi with NDV. GAPDH was used as a control. (D–G) Treatment with the NSC23766 and IPA-3 had no effect on the adsorption and internalization of NDV. HD11 cells were pretreated with NSC23766 (D and E), IPA-3 (F and G), after which NDV adsorption and internalization assays were performed. Flow cytometry was used to analyze the MFI of DiOC-labelled NDV. The bars represent the means ± SD from three independent experiments (* P < 0.05; ** P < 0.01; *** P < 0.001; NS, no significant difference).

Article Snippet: IPA-3 , PAK1 activation inhibitor , MCE , HY-15663.

Techniques: Activity Assay, Activation Assay, Western Blot, Control, Phospho-proteomics, Adsorption, Flow Cytometry

Journal: Virology Journal

Article Title: Inhibitor analysis revealed that clathrin-mediated endocytosis is involed in cellular entry of type III grass carp reovirus

doi: 10.1186/s12985-018-0993-8

Figure Lengend Snippet: Variables from the questionnaire of the HBSC study analyzed in this study

Article Snippet: Inhibitors were prepared as follows: pistop2, dynasore, rottlerin, nystatin, wortmannin, bafilomycin A1, Latrunculin B, nocodazole, IPA-3, and amiloride were purchased from ApexBio (Houston, USA).

Techniques: Concentration Assay, Inhibition

Journal: Virology Journal

Article Title: Inhibitor analysis revealed that clathrin-mediated endocytosis is involed in cellular entry of type III grass carp reovirus

doi: 10.1186/s12985-018-0993-8

Figure Lengend Snippet: Effect of inhibitors on the production of progeny virus. CIK cells were treated with different inhibitors at the indicated concentrations and then infected with GCRV104 (MOI = 5) for 5 days. Uninternalized virions were removed at 1 hpi. Rt-PCR assay of virus yield in the supernatants. a Pistop2 (5 μM and 1 μM) and CPZ (10 μM, 5 μM, and 1 mM) inhibit GCRV104 infection. b Rottlerin (5 μM and 2 μM) and wortmannin (5 μM and 2 μM) inhibit GCRV104 infection. c and d Nystatin (15 μM, 3 μM), Methyl-β-cyclodextrin (1 mM, 0.5 mM, 0.1 mM), Latrunvulin B (0.5 μM, 0.25 μM, 0.05 μM), nocodazole (10 μM, 5 μM, 1 μM), IPA-3 (10 μM, 5 μM, 1 μM), Amiloride (10 μM, 5 μM, 1 μM), and Bafilomycin A1 (2 nM, 1 nM, 0.2 nM) were used for analysis different pathways. Asterisks represent a significant difference from the control (unpaired t-test, * P < 0.05 and ** P < 0.01)

Article Snippet: Inhibitors were prepared as follows: pistop2, dynasore, rottlerin, nystatin, wortmannin, bafilomycin A1, Latrunculin B, nocodazole, IPA-3, and amiloride were purchased from ApexBio (Houston, USA).

Techniques: Virus, Infection, Reverse Transcription Polymerase Chain Reaction, Control