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We found that in cells with decreased RACK1 expression levels there is an increase in the extent of M2 internalization with over 60% of receptors internalized after 30 minutes compared to almost 45% of receptors internalized in cells with normal levels of RACK1 expression (Fig. 5).
A similar mechanism has been reported for the internalization of the junction protein occludin that becomes ubiquitinated and internalized after stimulation of the cell with VEGF [ 67].
We found similar receptor expression levels at time 0 and a significant decrease in both the rate and extent of M2 internalization when RACK1 was overexpressed, with only about 40% of the receptors internalized after 60 minutes compared to almost 70% of receptors internalized in cells overexpressing M2 only.
The amount of internalized EGFR increased gradually over 5 20 min period with about 70% of EGFR being internalized after 20 min of stimulation.
The results are expressed as a percentage of the total radioactivity internalized after 1 h of incubation.
The toxin is internalized after binding a high-affinity receptor, synaptic vesicle protein 2 (SV2), which is exposed in the cell membrane during the exocytosis process.
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It is interesting to note that pansomatostatin-like radioligands failing to internalize after binding to sst2 in vivo indeed showed poor uptake in sst2 + tissues in mouse models [31, 32].
We next evaluated by confocal microscopy whether the FITC-conjugated CPE peptide is able to internalize after binding to claudin-3 and claudin-4 receptors on the surface of chemotherapy-resistant ovarian cancer cells.
In the present study, we have investigated whether disruption of the rotomer toggle switch by mutation of this residue to phenylalanine (W243F) affects the receptor's ability to internalize after agonist challenge.
We used confocal microscopy to evaluate whether the FITC-conjugated-C-CPE 290-319 peptide was able to internalize after binding to claudin-3 and claudin-4 receptors on the surface of chemotherapy-resistant ovarian cancer cells.
Sixty-four other cells internalize after the endoderm precursors, leading to roughly half of the embryonic cells ending up in the interior of the embryo (S ulston et al. 1983; N ance and P riess 2002; H arrell and G oldstein 2011).
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