Figure 8

PCA of the DNA flowcytometer data. Data from FCM analysis of propidium iodide (PI) stained cells from each treatment were processed in Cyflogic FCM analysis package according to the following criteria: cells were gated on a window in the pulsetime PI intensity dot plot to exclude cell aggregates. The gated signals were represented as histograms: intensity, pulse time, forward scatter, sideward scatter. In the intensity plot, 4 windows were defined: DNA content below G1 phase of the cell cycle for apoptotic cells (Below G1), the window indicating G0/G1 phase (G1), a window for S-phase (S) and a window for G2/M phase (G2). In each window, the number of cells was expressed as % of total cell count in de gating window. The eigenvectors computed from these parameters are indicated on the plot by the small squares. Each dot is a separate treatment, indicated by the treatment code: C: control; Pl: P. linteus polysaccharide extract; Pl + HS: P. linteus polysaccharide extract complemented with human serum; HS: human serum; PMA: PMA treatment; Abi: A. bisporus polysaccharide extract; Abl: A. brasiliensis polysaccharide extract; codes are followed by the time at which cells were sampled (h) after begin of the treatment. A minimum of 1500 cells was counted in the gating window. Similar patterns are grouped and indicated with their most prominent feature in italics: Control for the control and HS24 treatment, Stimulated for the PMA and Agaricus polysaccharide extract treatments at 24 h, 48 dividing for PMA and A. bisporus and A. brasiliensis polysaccharide extracts treatments at 48 h, 48 inhibited for the cells arrested in G1 and P. linteus for the P. linteus polysaccharide extract treatments, resulting in sub G1 DNA levels due to fragmenting nuclei and dead cells.