Effect of Rottlerin on acid secretion. Human CD14+ monocytes were isolated from blood and seeded in 96 well plates and cultured in the presence of 25 ng/ml M-CSF and RANKL for 12 days. The mature human osteoclasts were incubated for 45 minutes with acridine orange alone or in combination with different concentrations of Rottlerin or Bafilomycin A1 (200 nM) as a positive control. All the results are presented as % of Bafilomycin. A) Acridine Orange pictures showing the effect of Rottlerin. B) Acridine Orange readings of the green wavelength (Excitation 492 nm - emission 535 nm) was measured using a SpectraMax M5 showing the effect of Rottlerin. The results are representative of three individual experiments performed in quadruplicates. The asterisks indicate significant differences between vehicle and inhibitor treated conditions.