Then the co-cultured cells were rigorously washed twice with phosphate buffered saline (PBS) to remove bacteria in suspension, trypsinized to detach mammalian host cells for flow cytometry , fixed with PBS supplemented with 1% formaldehyde, 0.33% BGS and 0.001% of sodium pyruvate and assayed for their GFP signals by flow cytome-try (FACSCanto II, Becton Dickinson).

(B) Flow cytometry of bacterial GFP associated with mammalian cells.

(D) Flow cytometry detection of surface Bl-integrins.

Flow cytometry data for GFP expression.

After co-culture of increasing amounts of GFP-expressing E. coli harboring a plasmid permitting arabinose-inducible control of invasin (pBACr-Aralnv) with HeLa cells, flow cytometry was conducted to obtain data shown in Fig 3A.

To simplify the data for comparison, a centroid and mean level of Bl-integrins at a given fraction of infected host cells were computationally estimated from each flow cytometry sample at different MOIs (Fig 3B).

This property was consistent with flow cytometry measurements (Fig 1B): At intermediate bacterial concentrations, a bimodal distribution of GFP fluorescence arises where within a single population there exist both uninfected cells (i.e.

Analysis of cell phenotype and cytokine secretion by flow cytometry

Following blocking of Fc receptors with antibodies, cells were labeled with CD 14 (PerCP-Cy5.5), CD1 1b (FITC), TIE-2 (Alexa 647), VEGFR-l (PE), TGFBR-l (Pacific Blue), TNF-Rl (Pacific Orange), CXCR4-, CCR5-, oc5l31-biotinylated specific antibodies (followed by strepta-vidin-Marina Blue) and analysed by flow cytometry using a Facs LSRII (BD Biosciences) equipped with a 610/20 nm filter on the violet detector.

TEM release of angiogenic factors measured by flow cytometry .

First, changes in TEM phenotype were evaluated by flow cytometry 36h post treatment.

TEM phenotype, cytokine secretion and pro-angiogenic activity were assessed by flow cytometry and in vivo or in vitro vascularization assay, respectively.

TEM) cell populations were measured by flow cytometry 36 hours posttreatment and displayed as mean logz ratios relative to untreated cells.

We characterized by flow cytometry the phenotype of TEM from patient peripheral blood and freshly dissociated tumor specimens obtained at time of surgery (see Material and Methods).

Based on our immunostaining and flow cytometry protocol we observed that TEM did not constitute a distinct subset of monocytes.

Expression levels of receptors and integrin measured by flow cytometry at the surface of TEM.

One of the most common techniques to reconstruct traction force based on TFM data is Fourier Transform Traction Cytometry (FTTC) [9,10].

From the displacement data, Fourier transform traction cytometry (FTTC) [9] was then used to estimate traction stress [10].

(C) Reconstruction of the traction forces with regularized Fourier Transform Traction Cytometry depends on the choice of a regularization parameter.