Fig. 3.

N- and O-glycan processing mediate early steps in differentiation. (A) WT Y101 hTERT-MSCs were pre-treated with the indicated inhibitors for 48 h before differentiation was induced in osteogenic medium in the absence of inhibitors for 21 days (inhibitors were included for the full 21 days for comparison where indicated as ‘continuous’). ALP and von Kossa staining was performed on the indicated days. The images in the last row show Cog4KD cells rather than WT. Scale bar: 100 µm (top three rows), 5 mm (bottom five rows). (B) Real-time qPCR experiments performed as in Fig. 2C with Y101 cells cultured in osteogenic medium in the continuous presence of kifunensine following a 48 h pre-treatment or pre-treated with kifunensine for 48 h followed by incubation in medium without kifunensine. The +kif data are repeated from Fig. 2 for clarity. Data represent averages of mean±s.e.m. of three technical repeats each for two independent biological replicates. *P<0.05, ***P<0.001 (compared to the control samples), all other changes are non-significant.