Figure 5.

Synapsin I expression in ASC cultures. The different conditions were investigated by immunocytochemistry and flow cytometry after 1 (A) and 7 (B) days of culture. In each row, microphotographs are followed by flow cytometry histograms (fourth column) in the same conditions. Quantitative immunofluorescence data (ImageJ) and flow cytometry mean fluorescence intensity (MFI) are also reported in panels (C,D), respectively. A virtually absent Synapsin I expression is detected when ASCs are grown in their basal medium, on both day 1 and 7, regardless of the presence of melatonin (first column). On day 1, a significant increase is observed when glial CM are used (second and third column), with no evident effects when melatonin was also present. Comparable increases were induced on day 7 by CM treatments; a greater increase was obtained using the combination melatonin/SC-CM. Blue fluorescence counterstaining indicates cell nuclei. Scale bars: 50 µm. Quantitative data reported in the histograms (panels C,D) confirm Synapsin I increases following glial CM. A melatonin-induced increase on day 7 in combination with SC-CM is also confirmed. Each plot in panel (C) summarizes a total of 60 measurements. * p < 0.05 melatonin presence vs. corresponding melatonin absence; # p < 0.05 day 7 vs. day 1; § p < 0.05 OEC-CM vs. SC-CM with or without melatonin.