Fig. 5
From: A novel method to efficiently differentiate human osteoclasts from blood-derived monocytes
Osteoclasts differentiated from Teflon bag-derived macrophages express typical osteoclast markers. A Flow cytometry: Expression of surface marker proteins (CD45, CD14, CD51) was assessed on monocytes, macrophage and bag-derived osteoclasts on n > 3 donors. Shown is one representative histogram per staining. Blue: antibody-of-interest, grey: isotype control. B Expression of known osteoclast marker genes was measured by qPCR (line at median). C + D Expression of selected osteoclast marker proteins was analyzed via western blotting in whole cell lysates (20 µg) of macrophages and osteoclasts (C). Signals were quantified by densitometry and normalized to Tubulin expression (line at median) (D). Outliers were identified and excluded according to the ROUT method (Q = 1%). E Expression of early- (DCSTAMP, NFATC1, CA2) and late-stage (ATP6V0A1, MMP9, MMP14) osteoclast markers was measured by qPCR in osteoclasts differentiated in cell culture plates or from Teflon bag-derived macrophages (line at median)