Fig. 1

The pattern of NTPDases expression in human BM-MSCs. Panel A presents representative images of the immunocytochemical detection of NTPDase1, −2, −3, −8 and ecto-5′-nucleotidase (Ecto 5'NTase/CD73) in cultured BM-MSCs (first subculture) obtained from a young female and a Pm woman. Shown is the time-related immunoreactivity (green) detected by confocal microscopy in BM-MSCs allowed to grow for 7 and 21 days in an osteogenic-inducing medium. Blue dots represent nuclei stained with DAPI. Experiments were performed in parallel keeping unaltered the settings of the confocal microscope throughout the documentation procedure (see Materials and Methods). The scale bar is 50 µm. In panel B, shown are immunofluorescence intensity graphs computed from confocal microscopy images acquired as in panel A. Ordinates represent fluorescence intensity per cell (arbitrary units, a.u.) of enzymes immunoreactivity as a function of the number of days in culture (day 7 and 21). Regions of interest (ROIs, corresponding to individual BM-MSCs) were manually outlined and the average intensity of the pixels inside this area was calculated. Background fluorescence was estimated from outlined regions (with no cells) and subtracted from all monitored ROIs. A total of 58–234 cells were analysed from three young females (21 ± 7 years old) and six Pm women (63 ± 4 years old). *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 (one-way ANOVA with Sidák’s multiple comparison test, single pooled variance) represent significant differences