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To assess the degree of morphological disruption, we examined the pattern of actin and ElaV expression in each eye epithelium.
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The severity of morphological disruption morphology was scored according to an arbitrary index with 0 being normal and 4 being severely disrupted (Table 1).
Because GMR-Gal4 drives expression initially in third instar larvae, we assessed morphological disruption at this stage.
Using this metric, we demonstrated a statistically significant rescue of CagA morphological disruption by co-expression of sqhA21 (Fig. 3G).
In the present study, the mitochondria in F20 group showed morphological disruption whereby the sizes were increased, the cristae were disrupted, and the matrixes were hypodense compared to C group.
Taking advantage of the temperature dependency of the Gal4-UAS system, we asked whether raising temperature would enhance CagAEPISA's ability to induce morphological disruption.
Our work in the eye epithelium argued that CagA induces morphological disruption by activating the Rho/MLC pathway.
We showed that the reduced potency of the CagAEPISA mutant could not be explained by a role for SHP-2/Csw in the morphological disruption.
However, morphological disruption was greatly reduced when CagA was co-expressed with a single copy of the inactivating sqhA21 mutation (Fig. 3C).
On its own, CswDN did not cause morphological disruption at larval stages, but did cause a mild rough eye phenotype in adults (not shown).
Furthermore, we showed that the EPIYA motifs of CagA are necessary for proper apical targeting in the polarized retinal epithelial cells, and loss of these motifs renders CagA less potent in inducing morphological disruption in the epithelium.
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