Figure 4
Validation of proteomics data by flow cytometry. (a) Fresh human monocytes from three different donors were analysed by flow cytometry with antibodies specific to the indicated protein, in addition to anti-CD86, anti-CD14 and anti-CD16 to distinguish each monocyte subset. Corresponding profiles as determined by MS are shown in the lower panel. For both types of analyses, data were normalised to a maximum of one then averaged across the three replicates. Data are shown as mean +/− SEM. For proteomic data, a Benjamini-Hochberg-corrected two-tailed t-test was used to estimate p-values. For flow cytometry data, a two-tailed t-test was used to estimate p-values. *p < 0.05; **p < 0.005. (b) Representative contour plots from a single donor of flow cytometry data for each illustrated marker coloured by cell subset. FSC-A (Forward Scatter Area.) was used as an intrinsic description of cell size (which would not be expected to change between subsets) to separate the points on a second axis. This representation was advantageous compared to the alternative of histograms as it more clearly visually represents the distribution of data points with less confounding by absolute cell number. The relative proportion of each monocyte subset is different in peripheral blood, with classical monocytes predominating.
