Figure 3

Increase in integrin β5 and decreases in SERPINE2, Perforin 1, and WARS are validated at the protein level in osteoclasts from osteopetrotic patients vs those from healthy donors. (a) Western blot of proteins extracted from two osteopetrotic patients (mutations G215R and R767W) and two healthy donor osteoclast cultures showing greater expression of the ITGB5, no difference in CES1 expression, and lower expression of SERPINE2 in osteopetrotic osteoclasts vs healthy donor osteoclasts. The expression is expressed as the ratio of the protein expression over GAPDH expression. (b) Western blot of proteins extracted from two osteopetrotic patients and two healthy donor osteoclast cultures showing lower expression of WARS and PRF1. The expression is expressed as the ratio of the protein expression over GAPDH expression. (c) Immunofluorescence was performed on all the osteopetrotic and healthy donors osteoclasts. The immunostainings shown here are representative of those observed for all the osteoclast cultures performed. It shows the cytoplasmic expression of ITGB3, ITGB5, SERPINE2, PRF1, and WARS (revealed by a secondary antibody coupled with Alexa488) (first column) in human osteoclasts in comparison with the phalloidin localization (Texas Red) (second column), nuclei stained with 4′,6-diamidino-2-phenylindole (DAPI) (third column), and a merge image (fourth column) (scale bar: 50 μm).