Cells (1×106) from each group were washed twice with precooled PBS and subjected to flow cytometry using a Cell Cycle Assay kit (BD Biosciences, Franklin Lakes, NJ, USA) according to the manufacturer's protocol. Cell cycle distribution was analyzed using ModFit software (version 5.0; Verity Software House, Topsham, ME, USA).
Following treatment with sera from healthy subjects, patients with sepsis or patients with septic shock for 24 h, cells (1×106) from each group were washed with precooled PBS twice and subjected to flow cytometry using an Annexin V fluorescein (FITC) Apoptosis Detection kit I (BD Biosciences; included PI staining materials) according to the manufacturers protocol. Cells with Annexin V-positive staining were early apoptotic cells, those with propidium iodide (PI)-positive values were necrotic and double-positive values were cells in late apoptosis. ModFit software (version 5.0; Verity Software House, Inc., Topsham, ME, USA) was used for the analysis of results.
Following treatment with sera from healthy subjects, patients with sepsis or patients with septic shock for 24 h, cells (1×106) from each group were washed with precooled PBS twice and subjected to flow cytometry using an Annexin V fluorescein (FITC) Apoptosis Detection kit I (BD Biosciences; included PI staining materials) according to the manufacturers protocol. Cells with Annexin V-positive staining were early apoptotic cells, those with propidium iodide (PI)-positive values were necrotic and double-positive values were cells in late apoptosis. ModFit software (version 5.0; Verity Software House, Inc., Topsham, ME, USA) was used for the analysis of results.