The PI3K pathway regulates endochondral bone growth through control of hypertrophic chondrocyte differentiation-7

2 or DMSO as vehicle control. They were stained after 6, 9 and 12 days, respectively for different chondrocyte differentiation markers: Alcian blue for glycosaminoglycans, Alkaline phosphatase activity and Alizarin red for the calcium content. The intensity of these markers is reduced in the LY294002 treated cultures. (B) After 9 and 12 days respectively the Alcian blue content of the micromasses was spectrophotometrically measured at 620 nm, after extraction with 6 M Guanidine hydrochloride. We noticed decreased absorbance levels for the LY294002 treated cultures. (C) Measurements of Hoechst fluorescence intensity (excitation/emission: 350/450 nm) showed no significant difference in the DNA content between the LY294002 and DMSO treated micromass cultures. (D, E) RNA was isolated from the micromass cultures after 6 and 9 days of culture and real time analysis was performed. The relative transcript levels for and were decreased in the LY294002 treated micromasses compared to the vehicle control.Copyright information:Taken from "The PI3K pathway regulates endochondral bone growth through control of hypertrophic chondrocyte differentiation"http://www.biomedcentral.com/1471-213X/8/40BMC Developmental Biology 2008;8():40-40.Published online 11 Apr 2008PMCID:PMC2329617.