Figure 5

SR-A inhibition rescues some of the oxLDL mediated effects in osteoclasts. (A) Increased dissolution of hydroxyapatite coated plates when SR-A is inhibited. Resorptive pits were visualized using a bright field microscope and quantified by ImageJ. Magnification x10 scale bar 1 mm. (B) SR-A inhibition did not improve the resorptive activity of oxLDL treated cells on bone slices. Magnification x10, scale bar 1.5 mm. (C) Conditioned media from B was collected and analyzed for the presence of collagen type 1 fragment using a colorimetric ELISA method. (D) CatK activity by fluorometric assay using synthetic substrate in oxLDL treated cells with and without SR-A inhibition. (E) mRNA expression of MMP-9 with/without oxLDL and SR-A inhibition. (F) Conditioned media from control and oxLDL treated osteoclasts were subjected to gelatin zymography with SR-A, CD36 and TLR4 inhibitions. (G) Conditioned media were subjected to western blot analysis and probed with the CatK antibody after SR-A, CD36 and TLR4 inhibitions. (H) Conditioned media from control and oxLDL treated osteoclasts grown on bone slices were subjected to western blot analysis and probed with the CatK antibody. (I) Cells were lysed at day 3 and equal amount of cell lysates were western blotted for CatK. Only inhibition of SR-A increased the secretion of CatK, hence decreasing CatK content in cell lysates. Images from both zymography and western blot were captured using the VersaDoc‐3000 (Bio‐Rad Laboratories, Muenchen, Germany).