Figure 4
microRNA-mediated repression of UGCG in CHO cells. (A) Enzymatic activity of UGCG was determined by conversion of NBD-Cer to NBD-GlcCer on TLC. Left, Lane 1, Standard lipids; Lane 2, lipids extracted from control cells; Lane 3, lipids extracted from UGCG depleted cells. Right, Quantification of lipids was shown. (B) Total GSLs extracted from cells were subjected to TLC followed by staining with orcinol/sulfuric acid. Lane 1, GM3 standard; Lane 2, GSLs extracted from EC2-1H9 cells; Lane 3, GSLs extracted from EC2-1H9-miUGCG cells. n = 3. (C) Quantification of total GSLs. n = 3, **P < 0.01 (Two-tailed Student’s t-test). (D) Cell proliferation was determined for 6 days of culture. Closed circles, EC2-1H9 cells; Open circles, EC2-1H9-miUGCG. n = 3. Data are presented as means ± S.E.M.
