Figure 3. Total sGAG and C4S increased by hypoxia in BEC and NCM460 cells.

A. In the BEC and the NCM460 cells, the total sGAG increased significantly following 10% O₂×24 h (p = 0.002, p = 0.013 respectively, unpaired t-test, two tailed), consistent with an effect on ARSB function. B. The majority of the increase in sGAG level following 10% O₂×24 h was attributable to the increases in C4S in the BEC and the NCM460 cells (p<0.001, p = 0.002, respectively). C, D. Highly significant inverse correlations were present between the sGAG content and the ARSB activity, using measurements of sGAG and ARSB at baseline, following ARSB knockdown by siRNA, following hypoxia (10% O₂×24 h), and following ARSB overexpression in the BEC (r = −0.96) and NCM460 cells (r = −0.999). E, F. Similar highly significant inverse correlations were present between the C4S content and the ARSB activity in the BEC (r = −0.95) and NCM460 cells (r = −0.99). [sGAG = sulfated glycosaminoglycan; BEC = bronchial epithelial cells; C4S = chondroitin-4-sulfate].