Figure 3

BDNF and NGF are weakly expressed in BM-MSCs. Cultured cells were treated with C5aR-agonist or antagonist for 48 h and then stimulated with LPS for 1 h, and fixed and stained accordingly. Immunofluorescence double staining was used to analyze the expression of BDNF and NGF. Anti-BDNF (green) and anti-NGF (red) were found weakly co-localized in the cytoplasm of BM-MSCs in C5aR-agonist treated cells after 48 h of treatment. Staining was observed in C5aR-agonist and LPS + C5aR-agonist treated cells. For each condition, negative controls, performed by replacement of the BDNF and NGF primary antibodies and secondary antibody used to detect BDNF was Alexa-488 (green) and NGF through Alexa-594 (red); nuclei were counterstained with DAPI (blue). Scale bar: 100 µm.