As DDX21 is localized in the nucleus [10] and translation and
replication of DENV occur in the cytoplasm, we examined whether
DENV infection might modify the nuclear localization of DDX21.We
used confocal laser scanning microscopy and immunofluorescence
to determine the subcellular distribution of DDX21 during DENV
infection of A549 cells, with an antibody specific to dsRNA that has
been reported previously as the marker for DENV infection [22].We
observed that the DDX21 signal was reduced in DENV infected cells
(dsRNA signal) and could barely be detected in the cytoplasm of
either DENV-infected or control cells (Fig. 2A). Additionally, the
protein levels of DDX21 from nucleus and cytoplasm of DENVinfected
or mock infected cells were evaluated by immunoblotting.
The results showed that the amount of DDX21 in the cytoplasm
of infected cells was slightly increased compared to
uninfected cells (Fig. 2B). However, the amount of DDX21 detected
in the nucleus of infected cells was reduced to one third compared
to the uninfected cells (Fig. 2C). These results, together with the
result in Fig. 1, suggest that DENV infection causes partially relocation
of DDX21 from the nucleus to the cytoplasm where the
protein is presumably degraded as we did not observe significant
accumulation of DDX21 in the cytoplasm compared to the amount
reduced in the nucleus.
As DDX21 is localized in the nucleus [10] and translation andreplication of DENV occur in the cytoplasm, we examined whetherDENV infection might modify the nuclear localization of DDX21.Weused confocal laser scanning microscopy and immunofluorescenceto determine the subcellular distribution of DDX21 during DENVinfection of A549 cells, with an antibody specific to dsRNA that hasbeen reported previously as the marker for DENV infection [22].Weobserved that the DDX21 signal was reduced in DENV infected cells(dsRNA signal) and could barely be detected in the cytoplasm ofeither DENV-infected or control cells (Fig. 2A). Additionally, theprotein levels of DDX21 from nucleus and cytoplasm of DENVinfectedor mock infected cells were evaluated by immunoblotting.The results showed that the amount of DDX21 in the cytoplasmof infected cells was slightly increased compared touninfected cells (Fig. 2B). However, the amount of DDX21 detectedin the nucleus of infected cells was reduced to one third comparedto the uninfected cells (Fig. 2C). These results, together with theresult in Fig. 1, suggest that DENV infection causes partially relocationof DDX21 from the nucleus to the cytoplasm where theprotein is presumably degraded as we did not observe significantaccumulation of DDX21 in the cytoplasm compared to the amountreduced in the nucleus.
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