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miglustat  (MedChemExpress)
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MedChemExpress miglustat
Glycosylation inhibitors reduce PEDV replication in Vero cells. ( A ) Schematic of the key steps in the N-glycosylation pathway targeted in this study. ( B ) Viability of Vero cells treated with the indicated concentrations of glycosylation inhibitors (tunicamycin at 5 and 50nM, NGI-1 at 1 and 5µM, <t>miglustat</t> at 5 and 10µM, and celgosivir at 40 and 80µM) for 24hours. ( C–F ) Vero cells pretreated for 1hour with inhibitors and infected or not (uninfected) with PEDV (MOI of 0.01, 20hours) in a fresh medium containing inhibitors. ( C ) Relative PEDV N mRNA levels determined by RT-qPCR and expressed relative to that in dimethyl sulfoxide (DMSO)-treated cells. GAPDH was used as the internal loading control. ( D ) Western blot of N-protein in cells infected with PEDV and treated with the indicated concentrations of glycosylation inhibitors or DMSO. ( E ) Immunofluorescence assay (IFA) images of Vero cells infected with PEDV and treated with indicated inhibitors. Viral N-protein is shown in green, and the nuclei are blue. Scale bars, 500µm. ( F ) Culture supernatants were collected for viral titration. No infectious virus particles were detected in the supernatant of cells treated with tunicamycin at a concentration of 50nM. The data are presented as the means ± SDs. The asterisks indicate significant differences (* P < 0.05; ** P < 0.01).
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Glycosylation inhibitors reduce PEDV replication in Vero cells. ( A ) Schematic of the key steps in the N-glycosylation pathway targeted in this study. ( B ) Viability of Vero cells treated with the indicated concentrations of glycosylation inhibitors (tunicamycin at 5 and 50nM, NGI-1 at 1 and 5µM, miglustat at 5 and 10µM, and celgosivir at 40 and 80µM) for 24hours. ( C–F ) Vero cells pretreated for 1hour with inhibitors and infected or not (uninfected) with PEDV (MOI of 0.01, 20hours) in a fresh medium containing inhibitors. ( C ) Relative PEDV N mRNA levels determined by RT-qPCR and expressed relative to that in dimethyl sulfoxide (DMSO)-treated cells. GAPDH was used as the internal loading control. ( D ) Western blot of N-protein in cells infected with PEDV and treated with the indicated concentrations of glycosylation inhibitors or DMSO. ( E ) Immunofluorescence assay (IFA) images of Vero cells infected with PEDV and treated with indicated inhibitors. Viral N-protein is shown in green, and the nuclei are blue. Scale bars, 500µm. ( F ) Culture supernatants were collected for viral titration. No infectious virus particles were detected in the supernatant of cells treated with tunicamycin at a concentration of 50nM. The data are presented as the means ± SDs. The asterisks indicate significant differences (* P < 0.05; ** P < 0.01).

Journal: Journal of Virology

Article Title: STT3B promotes porcine epidemic diarrhea virus replication by regulating N-glycosylation of PEDV S protein

doi: 10.1128/jvi.00018-25

Figure Lengend Snippet: Glycosylation inhibitors reduce PEDV replication in Vero cells. ( A ) Schematic of the key steps in the N-glycosylation pathway targeted in this study. ( B ) Viability of Vero cells treated with the indicated concentrations of glycosylation inhibitors (tunicamycin at 5 and 50nM, NGI-1 at 1 and 5µM, miglustat at 5 and 10µM, and celgosivir at 40 and 80µM) for 24hours. ( C–F ) Vero cells pretreated for 1hour with inhibitors and infected or not (uninfected) with PEDV (MOI of 0.01, 20hours) in a fresh medium containing inhibitors. ( C ) Relative PEDV N mRNA levels determined by RT-qPCR and expressed relative to that in dimethyl sulfoxide (DMSO)-treated cells. GAPDH was used as the internal loading control. ( D ) Western blot of N-protein in cells infected with PEDV and treated with the indicated concentrations of glycosylation inhibitors or DMSO. ( E ) Immunofluorescence assay (IFA) images of Vero cells infected with PEDV and treated with indicated inhibitors. Viral N-protein is shown in green, and the nuclei are blue. Scale bars, 500µm. ( F ) Culture supernatants were collected for viral titration. No infectious virus particles were detected in the supernatant of cells treated with tunicamycin at a concentration of 50nM. The data are presented as the means ± SDs. The asterisks indicate significant differences (* P < 0.05; ** P < 0.01).

Article Snippet: Tunicamycin, NGI-1, miglustat, and celgosivir were purchased from MCE (USA).

Techniques: Infection, Quantitative RT-PCR, Control, Western Blot, Titration, Virus, Concentration Assay