ma 104  (ATCC)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Integrated phenotypic screening and chemical proteomics identifies ETF1 ligands that modulate viral translation and replication

doi: 10.1073/pnas.2524108123

Figure Lengend Snippet: Photo-stereoprobes inhibit additional viruses that use PRF. ( A ) Effect of active stereoprobe WX-02-31 and control stereoprobes on HIV-1 pseudovirus production. ( Top ) Schematic of the HIV-1 ribosomal frameshift sequence. ( Bottom ) HEK293T cells expressing the HIV-1 NL4-3 luciferase vector were treated with the indicated photo-stereoprobes (10 µM) and assessed for luciferase activity after 24 h. Data are average values ± SD for 2 to 5 independent experiments that each have three technical replicates. ( B ) Effect of active stereoprobe WX-02-31 and control stereoprobes on Sindbis virus (SINV) infection. ( Top ) Schematic of the SINV ribosomal frameshift sequence. ( Bottom ) HeLa cells were pretreated with indicated photo-stereoprobes (10 µM, 3 h), inoculated with SINV (MOI 1) in the presence of compounds, and viral supernatant titers were assessed after 48 hpi. Data are average values ± SD for three independent experiments that each have 2 to 3 technical replicates. ( C ) Effect of active stereoprobe WX-02-31 and control stereoprobes on porcine reproductive and respiratory syndrome virus (PRRSV) infection. ( Top ) Schematic of the PRRSV ribosomal frameshift sequence. ( Bottom ) MA-104 cells were pretreated with indicated photo-stereoprobes (10 µM, 3 h), inoculated with PRRSV-GFP (MOI 0.1) in the presence of compounds, and viral antigen levels were assessed by GFP expression after 24 hpi. Data are representative of two independent experiments that each have three technical replicates. ( D ) Effect of active stereoprobe WX-02-31 and control stereoprobes on lymphocytic choriomeningitis virus (LCMV) infection. ( Top ) Schematic of the LCMV genome. ( Bottom ) HeLa cells were pretreated with indicated photo-stereoprobes (10 µM, 3 h), inoculated with LCMV (clone 13, MOI 0.01) in the presence of compounds and viral supernatant titers were assessed after 48 hpi. Data are average values ± SD for two independent experiments that each have 2 to 3 technical replicates. ( E ) Effect of active stereoprobe WX-02-31 and control stereoprobes on influenza A virus (IAV) infection. ( Top ) Schematic of the IAV genome. ( Bottom ) Calu-3 cells were pretreated with indicated photo-stereoprobes (20 µM, 3 h), inoculated with IAV (MOI 0.005) in the presence of compounds, and viral antigen levels were assessed by flow cytometry after 48 hpi. Data are the average values ± SD for two independent experiments that each have three technical replicates. For ( A – C ), two-tailed unpaired t test comparing WX-02-31 versus DMSO or WX-02-51 groups: * P < 0.05, **** P < 0.0001.

Article Snippet: Calu-3 (HTB-55), Vero E6 (CRL-1586), HEK-293T (CRL-3216), HeLa (CCL-2), NIH-3T3 (CRL-1658), MA-104 (CRL-2378.1), and MDCK (CCL-34) cells were obtained from the American Type Culture Collection (ATCC).

Techniques: Control, Sequencing, Expressing, Luciferase, Plasmid Preparation, Activity Assay, Virus, Infection, Flow Cytometry, Two Tailed Test

Journal: Journal of Virology

Article Title: A small-molecule HSP90 inhibitor, NVP-HSP990, alleviates rotavirus infection

doi: 10.1128/jvi.01883-25

Figure Lengend Snippet: Cytotoxicity and inhibitory effects on RV replication of different HSP90 inhibitors. ( A ) Plots for cell viability of MA104, Caco-2, and HT-29 cells after treatment with NVP-HSP990, GA, or 17-AAG at indicated concentrations for 24 h. Cell viability was tested using the CCK-8 assay. ( B ) Plots for RV (Wa and SA11 strains) inhibition in MA104, Caco-2, and HT-29 cells after treatment with NVP-HSP990, GA, or 17-AAG at indicated concentrations for 24 h. RV replication was tested by PFA, and IC 50 values are indicated at the top of each plot. The experiments were performed in triplicate, and the data are presented as mean ± SEM and are representative of four ( A ) and two ( B ) independent experiments. ns, not significant; * P < 0.05, ** P < 0.01, **** P < 0.0001 (two-way ANOVA).

Article Snippet: Rhesus monkey embryo kidney cell line MA104 cells (ATCC: CRL-2378.1) were provided by Dr. Elschner (Friedrich-Loeffler-Institute).

Techniques: CCK-8 Assay, Inhibition

Journal: Journal of Virology

Article Title: A small-molecule HSP90 inhibitor, NVP-HSP990, alleviates rotavirus infection

doi: 10.1128/jvi.01883-25

Figure Lengend Snippet: NVP-HSP990 inhibited MAPK activation and facilitated expression of tight junction-associated proteins in intestinal cells. ( A ) MA104, Caco-2, and HT-29 cells were mock-infected with PBS or infected with RV Wa or SA11 strains (MOI = 3), followed by treatment of 100 nM HSP990 (+) or an equal volume of DMSO as a control (−) for 20 h. Then the infected cells were harvested for WB analysis of MAPK components. ( B ) Caco-2 cells were mock-infected with PBS, treated with 1 µM C16-PAF(C16), or infected with RV Wa or SA11 strains (MOI = 3), and then treated with 100 nM HSP990 (+) or DMSO as a control (−) for 20 h. The infected cells were harvested for WB analysis of MAPK components. ( C ) Caco-2 cells were mock-infected with PBS or infected with RV Wa or SA11 strains (MOI = 3), and then treated with 100 nM HSP990 (+) or DMSO as control (−) for 20 h. Then the infected cells were harvested for WB analysis of tight junction-associated proteins. Data are representative of three ( A ) and two ( B and C ) independent experiments.

Article Snippet: Rhesus monkey embryo kidney cell line MA104 cells (ATCC: CRL-2378.1) were provided by Dr. Elschner (Friedrich-Loeffler-Institute).

Techniques: Activation Assay, Expressing, Infection, Control