Expression of SLRPs in HTB94 chondrosarcoma cells. (A) Specific primers for each gene were used and normalized against GAPDH as the housekeeping gene. Mean Ct values are presented from 6 replicates (SEM, standard error of the mean). (B) Representative western blots showing the expression of lumican, decorin and biglycan in the culture medium of HTB94 chondrosarcoma cells. (C) Protein bands of the 3 secreted SLRPs were densitometrically measured utilizing the ImageJ program and expressed as arbitrary units. (D) Following protein extraction from HTB94 harvested cells, the protein sample was loaded in an acrylamide gel. An anti-lumican antibody was used to determine the expression of lumican in the cell extracts. The position of the nearest respective protein marker band is depicted to the right. (Ea) Lumican (green; anti-mouse Alexa Fluor 488) protein staining and (Eb) nuclear staining (using TO-PRO-3) were evaluated in culture; (Ec) superimposed image; (Ed) in the negative control, the primary antibody was omitted, but the anti-mouse secondary antibody was used. Slides were analyzed by confocal microscopy, and images were obtained at ×40 magnification. Results represent the average of 3 separate experiments. Representative plots are presented. Data are the means ± SEM. SLRPs, small leucine-rich proteoglycans.