Paszkowski Patrick
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View article: Timing of expression of VACV post-replicative and late genes.
Timing of expression of VACV post-replicative and late genes. Open
(A) EGFPcro BSC-40 cells were infected at a MOI = 5 with a virus encoding mCherry-tagged to the I1 protein (VACV-I1L-mCherry) and then tracked via live cell microscopy. These are stills taken from S5 Video. (B) mCherry-cro BSC-40 cells wer…
View article: Summary of different reporter protein expression kinetics.
Summary of different reporter protein expression kinetics. Open
The plot shows when a mCherry-cro signal is first detected relative to the time when a factory is first detected in that cell (tf). Each imaging experiment was repeated 3 times, and 4/10 fields in each experiment were analyzed in detail, t…
View article: Southern blot analysis of recombinants.
Southern blot analysis of recombinants. Open
BSC-40 cells were infected (or co-infected) with the different indicated viruses, and Southern blotted to detect recombinants. (A) The scheme used to collect samples for Southern blotting. Some DNA was extracted directly from virus-infecte…
View article: Recombination between pE/L-mCherry(t) and pE/L-mCherry-lacZ viruses.
Recombination between pE/L-mCherry(t) and pE/L-mCherry-lacZ viruses. Open
(A) The figure shows the two parent viruses, the predicted recombinants, and the HindIII fragments that should be detected by the pE/L oligonucleotide probe (blue bar). (B) Southern blot analysis of cells co-infected with the pE/L-mCherry(…
View article: Maintenance of factory boundaries at an early stage of co-infection.
Maintenance of factory boundaries at an early stage of co-infection. Open
BSC-40 cells were co-infected with pE/L-mCherry-cro and pE/L-EGFP-cro viruses (schematics, top) for 4 h and then fixed and stained with DAPI to also detect virus and cell DNA. The images shown here are taken from a single Z-stack showing c…
View article: Timing the appearance of early (I3) and late (A34) genes by western blotting.
Timing the appearance of early (I3) and late (A34) genes by western blotting. Open
EGFPcro BSC-40 cells were cultured in 60 mm dishes, infected with the indicated viruses [(A) VACV-pE/L-mCherry-cro; (B) VACV-pE/L-mCherry(t) + VACV-mCherry-cro], and then different dishes were harvested at the indicated times. Protein extr…
View article: Tracking the appearance of virus-encoded mCherry proteins.
Tracking the appearance of virus-encoded mCherry proteins. Open
(A) EGFPcro BSC-40 cells were infected at a MOI = 5 with VACV-pE/L-mCherry-cro, and the red and green fluorescence then tracked over time, collecting images 5 minutes apart, across 10 different fields (only a single representative field is…
View article: Large late viral factories enclose internal ER membranes.
Large late viral factories enclose internal ER membranes. Open
(A) BSC-40 cells were infected with VACV strain WR for 4 h (middle panel) or 8 h (third panel) and then fixed and stained to detect DNA (DAPI), the ER membrane marker calreticulin, and the viral I3 protein. The images in Panel A show a pro…
View article: Timing of intraviral and virus-by-plasmid recombination events.
Timing of intraviral and virus-by-plasmid recombination events. Open
(A) EGFPcro BSC-40 cells were infected with VACV-pE/L-mCherry(dup) at MOI = 0.5 to favor infections with a single particle. Two different cells are tracked here from the time of factory development: one infected by an actively recombining …
View article: Quantifying the recombinants formed in co-infected cells.
Quantifying the recombinants formed in co-infected cells. Open
(A) The panel shows the two recombinant viruses that could be formed in cells co-infected with pE/L-mCherry(t) and mCherry-cro viruses. Also shown are the diagnostic restriction fragments that would be produced following XhoI digestion and…
View article: Characterization of the recombinant VACV constructed for this study.
Characterization of the recombinant VACV constructed for this study. Open
(A) Four recombinant viruses were constructed encoding combinations of the mCherry and Cro genes with or without a synthetic early-late pox promoter (pE/L), all inserted into the TK locus of WR VACV. For convenience, these are shown in the…