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The presence of serum abrogated completely the binding to viable cells.
These results show that serum factors are involved in the prevention of C1q binding to viable cells and in the processing of late apoptotic/secondary necrotic cells promoting cell death progression toward apoptotic bodies.
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(A ) Detection of BODIPY-NaD1 binding to viable and permeabilized U937 cells by flow cytometry.
Purified C1q alone binds, albeit weaker, also to viable cells.
Results were referred to viable cells, discriminated from dead cells by 7 AAD staining (eBioscience, San Diego, CA).
The system has two stable steady states zero - corresponding to non-viable cells and the positive corresponding to viable cells.
Ig production was normalised to viable cell number.
The values of cell viability were calculated as percentages of viable cell numbers from bromoindirubin-treated cells to viable cell numbers from the DMSO-treated cells.
Moreover, we have previously shown that non-viable cell populations bind approximately 100 times more plg, masking changes in cell-surface binding on viable cells [ 27].
SfbI-TED binding to unfixed, viable, adherent cells was visualized by conjugating TED-iPT constructs via IPD complementation to GFP fused to sIPD (Zakeri et al., 2012).
Propidium iodide (Sigma-Aldrich) was added to cells to gate viable cells.
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