mers cov nucleoprotein np antibody  (Sino Biological)

Journal: Journal of Virology

Article Title: MERS-CoV and SARS-CoV-2 infection in diverse human lung organoid-derived cultures

doi: 10.1128/jvi.01098-25

Figure Lengend Snippet: Generation of donor-derived lung organoids (hLO), organoid-derived monolayers (hLOm), and ALI (hLO ALI) cultures. ( A ) Schematic of hLO generation and derivation of hLOm and hLO-ALI cultures, followed by infection with SARS-CoV-2 or MERS-CoV. Schematic created using BioRender. ( B ) Identification of lung cell markers. hLOm, hLO, and hLO ALI cells were fixed in 4% paraformaldehyde, permeabilized and stained with antibodies against acetylated tubulin (cilia), KRT5 (Basal Stem Cells, white arrows), Muc5AC (mucus and mucus producing cells, Goblet Cells), CC10 (Club Cells), ZO-1 (zona occludens-1, tight junctions), PDPN1 (podoplanin-1, AT-1 cells, white arrows), SFTPC (surfactant protein C, AT-2 cells), and DAPI (nuclei, blue). Cells were imaged on a confocal microscope using a 63× objective. Scale bars correspond to 50 µm. ( C ) Gene expression of lung markers FoxJ1 , TUBB4B , TJP1 , TP63 , KRT5 , NGRF , Muc5AC , SCGB1A1 , PDPN1 , AQP5 (aquaporin 5, AT-1 cells), SFTPC and SLC34A2 (AT-2 cells) in hLO, hLOm, and hLO ALI was determined by RT-qPCR assays. Samples were assayed in technical duplicates, dCt was normalized to GAPDH, and expression was calculated by 2 −dCt . The mean of three independent samples is presented, and error bars are standard error of the mean (SEM).

Article Snippet: MERS-CoV nucleoprotein N , Sino Biological , 40068-RP01-200.

Techniques: Derivative Assay, Infection, Staining, Microscopy, Gene Expression, Quantitative RT-PCR, Expressing

Journal: Journal of Virology

Article Title: MERS-CoV and SARS-CoV-2 infection in diverse human lung organoid-derived cultures

doi: 10.1128/jvi.01098-25

Figure Lengend Snippet: MERS-CoV infectivity in donor-derived lung organoids (hLO), organoid-derived monolayers (hLOm), and ALI (hLO ALI) cultures. ( A ) Brightfield and ( B ) confocal imaging of immunostained cells mock infected or infected with MERS-CoV at an MOI of 0.3 at 4 days post infection. Cells were fixed, permeabilized, and stained with antibodies against the MERS-CoV nucleoprotein ( N ) and DAPI (nuclei) and imaged at 63×. Scale bars correspond to 200 µm ( A ) and 50 µm ( B ). White arrows indicate infected cells and dashed lines and asterisks (yellow) delineate areas within the ALI in which ZO-1 was absent, and the culture integrity was disrupted ( B ). ( C ) Total RNA was collected at specified times to determine expression of a viral genomic region upstream the E gene (UpE). ( D ) MERS-CoV virus titers in supernatant of hLOm and apical and basolateral media of hLO ALI assessed using TCID 50 assay. Host gene expression of IFNβ ( E ), IRF7 ( F ), IFIT1 ( G ), STAT1 ( H ), MX1 ( I ), OAS1 ( J ), RSAD2 ( K ), TNF-α ( L ), IL6 ( M ), and IL1β ( N ) was evaluated by RT-qPCR. The mock infected samples for hLO ALI are the same as in . For RT-qPCR assays, samples were assayed in technical duplicates, dCt was normalized to GAPDH, and virus gene expression was calculated as 1/dCt. Host gene expression was calculated as fold-change relative to the mean of mock infected controls using the 2 −ddCt method. The mean of three independent samples is presented; error bars are standard error of the mean (SEM). Empty circles = mock; Purple circles = hLOm; Teal circles = hLO; Orange circles = hLO ALI. Statistical analysis was performed by one-way ANOVA ( C–M ) or Student’s t test ( N ). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: MERS-CoV nucleoprotein N , Sino Biological , 40068-RP01-200.

Techniques: Infection, Derivative Assay, Imaging, Staining, Expressing, Virus, Gene Expression, Quantitative RT-PCR