Journal: Nature communications

Article Title: A stochastic assembly model for Nipah virus revealed by super-resolution microscopy.

doi: 10.1038/s41467-018-05480-2

Figure Lengend Snippet: Fig. 1 Distribution of NiV-M in PK13 cells. PK13 cells expressing NiV-M were fixed, permeabilized, and labeled with goat anti-GFP and anti-goat Alexa Fluor 647 antibodies. a Schematic illustration of the imaging planes of a cell. b x–y cross section (100 nm thick in z) of a region through the middle (position 1 in a) of a representative cell showing the M clusters. Scale bar: 1 μm. c x–y cross section (100 nm thick in z) of a region at the plasma membrane (position 2 in a) of a representative cell showing larger M clusters. Scale bar: 1 μm. d x–y cross section of a region (position 3 in a) of a cell showing a representative dome-like structure formed by M. Scale bar: 1 μm. e z-stacks of the x–y cross section of the dome-like structure boxed in d. Scale bar: 0.1 μm. f 3D surface reconstructed by using the M localizations in e, with M localization density projected on the 3D surface. A higher brightness indicates a higher localization density. One representative cell image out of three independent experiments (n ≥30) is shown

Article Snippet: PK13 (pig kidney fibroblast, American Type Culture Collection; ATCC, CRL6489)13,34 and HeLa cells (ATCC, CCL-2) were grown at 37 °C and 5% CO2 in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum, 50 IU penicillin per ml, 50 μg streptomycin per ml, and 2 mM glutamine (Life Technologies).

Techniques: Expressing, Labeling, Imaging, Clinical Proteomics, Membrane

Journal: Nature communications

Article Title: A stochastic assembly model for Nipah virus revealed by super-resolution microscopy.

doi: 10.1038/s41467-018-05480-2

Figure Lengend Snippet: Fig. 2 Distribution of NiV envelope glycoproteins on the plasma membrane of PK13 cells. PK13 cells expressing NiV-F or/and -G were fixed at 24 h post transfection and immunostained. Without permeabilization, NiV-F was immunostained by using a mouse anti-FLAG primary antibody and -G a rabbit anti- HA primary antibody. For single-color SMLM, Alexa Fluor 647 secondary antibodies were used for detection. For dual-color SMLM, Alexa Fluor 647 secondary antibodies were used for detection of F, and Cy3B secondary antibodies for G. a, b x–y cross section (100 nm thick in z) at the dorsal surface (position 3 in Fig. 1a) of a representative cell expressing F (a) or G (b). The boxed region is enlarged to show the detailed distribution pattern. c Comparison analyses of the localization densities of the F (red) or G (green) at the cell body versus membrane protrusions. Each data point was calculated using an area of 0.2 × 0.2 μm2. All data were normalized to the mean of the cell body. d Hopkins’ index of the F and G localizations from n = 30 cells. Lines represent the mean value and SD. The sample size is indicated in the parentheses. The p values were determined by two-tailed, unpaired t-test with Welch correction. e x–y cross section (100 nm thick in z) of a region at the dorsal surface (position 3 in Fig. 1a) of a representative cell co-expressing F (red) and G (green) with a pixel size of 10 nm. Scale bars: 1 μm. f The distribution of the DoC values between F and G molecules. One representative cell image out of three independent experiments (n ≥30) is shown

Article Snippet: PK13 (pig kidney fibroblast, American Type Culture Collection; ATCC, CRL6489)13,34 and HeLa cells (ATCC, CCL-2) were grown at 37 °C and 5% CO2 in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum, 50 IU penicillin per ml, 50 μg streptomycin per ml, and 2 mM glutamine (Life Technologies).

Techniques: Clinical Proteomics, Membrane, Expressing, Transfection, Comparison, Two Tailed Test

Journal: Nature communications

Article Title: A stochastic assembly model for Nipah virus revealed by super-resolution microscopy.

doi: 10.1038/s41467-018-05480-2

Figure Lengend Snippet: Fig. 3 Presence of M does not affect the arrangement of the envelope glycoproteins’ clusters on the membrane. PK13 cells were co-transfected with NiV- M, -F, and -G and fixed at 24 h post transfection. Without permeabilization, F was immunostained using a mouse anti-FLAG primary antibody and an anti- mouse Cy3B secondary antibody, and G an anti-HA primary antibody and an anti-rabbit Alexa Fluor 647 secondary antibody. a, b x–y cross section (100 nm thick in z) of a representative cell shows the superimposition of a wide field image for M (blue) and the corresponding SMLM images of F (red) and G (green) on the cell body (a) and membrane protrusions (b). Scale bar: 1 μm. c x–y cross section (100 nm thick in z) of the M-positive sites (c1, 2) boxed in b. Scale bar: 0.1 μm. d 3D surface reconstruction of the dome-like structure in c1, with F (red) and G (green) localization densities projected on the surface. A higher brightness indicates a higher localization density. e Comparison of the localization densities of F (red) or G (green) on the M- positive and M-negative regions at the dorsal surface of the cell. f Comparison of the Hopkins’ indices of F (red) and G (green) in the M-positive and M- negative regions at the plasma membrane. All data in e and f were normalized to the mean of the M-negative regions of the same cell. Lines represent the mean value and SD. The sample size is indicated in the parentheses. The p values were determined by two-tailed, unpaired t-test with Welch correction. g The distribution of the DoC values between F and G molecules in the M-negative and M-positive regions. h VLPs produced in PK13 cells expressing M, F, and G were adhered to fibronectin-coated coverslips, fixed, and stained for NiV-F and G via tags described above. The z-stacks of the x–y cross section of a VLP show the super-imposition of a wide field image of M (blue) and the corresponding SMLM images of F (red) and G (green). Scale bar: 0.1 μm

Article Snippet: PK13 (pig kidney fibroblast, American Type Culture Collection; ATCC, CRL6489)13,34 and HeLa cells (ATCC, CCL-2) were grown at 37 °C and 5% CO2 in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum, 50 IU penicillin per ml, 50 μg streptomycin per ml, and 2 mM glutamine (Life Technologies).

Techniques: Membrane, Transfection, Comparison, Clinical Proteomics, Two Tailed Test, Produced, Expressing, Staining

Journal: eLife

Article Title: HA stabilization promotes replication and transmission of swine H1N1 gamma influenza viruses in ferrets

doi: 10.7554/eLife.56236

Figure Lengend Snippet:

Article Snippet: Cell line ( Sus scrofa ) , Swine testis (ST) fibroblast , ATCC , ATCC Cat# CRL-1746, RRID: CVCL_2204 , .

Techniques: Expressing, Recombinant, Software, Virus