The percentage of cells with eGFP-LC3 translocation into dots (a minimum of 100 cells/sample) was counted in eGFP-LC3 expressing cells fixed in 3.7% formaldehyde and 0.19% picric acid (vol/vol) applying Zeiss Axiovert 100 M Confocal Laser Scanning Microscope.
To quantify this translocation we performed dot blot analysis using a specific antibody against phosphorylated (Ser536) activated NF-κB p65.
The Bcl-2-associated transcription factor/BTF is found in the nucleus in dot like structures and its translocation depends on apoptotic signals [ 19].
However, in vitro, no translocation of quantum dots across primary rat alveolar epithelial cells was observed (Fazlollahi et al. 2011; Geys et al. 2009).
For cerium dioxide, quantum dots, silica, and titanium dioxide nanoparticles, different translocation rates are found in vitro and in vivo.
Vertically linked red-to-blue dots are associated with MPs specific to this translocation event between chromosomes 1 and 12 (Fig. 3a, i and ii).
(C ) Smad1 activation: Smad1 nuclear translocation in HUVECs was measured without flow (dotted line: average) or after 16 hr of flow in the gradient chamber (n = 6, Mean ± SEM, ANOVA: F = 13.47, p < 0.0001).
Calcium depletion of control cells caused rapid translocation of occludin from cell-cell junctions into intracellular dot-like aggregates (Figure 9, arrows), thus indicating breakdown and internalization of the AJC.
(B ) NF-κB activation: p65 nuclear translocation in HDLEC was measured either in no flow (dotted line: average) or after 16 hr of flow in the gradient chamber (n = 4, Mean ± SEM, ANOVA: F = 34.32, p < 0.0001).
(B ) NF-κB activation: p65 nuclear translocation in HUVEC was measured either in no flow (dotted line: average) or after 16 hr of flow in the gradient chamber (n = 6, Mean ± SEM, ANOVA: F = 10.97, p < 0.0001).
To determine if Glu residues contribute to recognition by the Icm/Dot system, we analysed a 27 amino acid carboxyl terminal fragment of SidM that allowed translocation of a CyaA reporter.
