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Although these mutants were internalized at a high rate, this increase in initial internalization is not sufficient to explain the hypercytotoxic phenotype.
Despite this, these results clearly indicate that this method can be accurately used not only to assess whether a bead has been internalized or not, but also to establish a rate of internalization by counting the number of beads that have been internalized at a given time (further details on the kinetics provided later in the Results).
Moreover, they were actively internalized at a temperature of 37°C but restricted to the cell membrane at 4°C.
While dead bacteria continue to be internalized at a high rate, the increased toxicity requires viable Francisella.
Although this is the first example of a LPS mutant in Francisella being internalized at a higher rate, enhanced host cell uptake of LPS mutants in other microbes has been shown.
Mutations in two other genes that are required for the addition of the O-antigen to the LPS including wzx and ΔwbtA were attenuated in mice [9,X.H. Lai and F. Heffron unpub data] but were not more cytotoxic to macrophages nor were they internalized at a higher rate (Figure 10B).
Similar(48)
The non-specifically bound dye is not always problematic; it is expected to be internalized at an efficiency similar to the internalization efficiency of the protein, and may or may not dissociate from the protein once internalized.
But the meme, at once personal and shared by millions, is a vector of almost unlimited value (as it has been by other names for millennia, as any scholar will tell you), and they are being created and internalized at an incredible rate.
The number of parent bacteria internalized at an input MOI of 10,000 was confirmed visually to be comparable to the mutant strains infected at a MOI of 100 (Figure 6B vs. Figure 4) and internalization of MFN245 at 10,000 MOI remained dependent upon actin polymerization (Figure 6B).
Ramos-CD21hi clone 1 internalized anti-CD19 at a similar low rate to ARH77 cells, while the more moderately expressing clone 3 internalized at an intermediate rate, in agreement with the immunofluorescence data.
As no burden sharing targets have been agreed it is hard to argue that burden sharing has been internalized at any level.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com