(A–B) Viability of MiaPaCa2 (A) and BxPC3 (B) empty vector (EV) and ST6Gal-I knockdown (ST6-KD) cells grown in CSC culture for two weeks. (C) Viability of SKOV3 EV, ST6-KD or ST6Gal-I overexpressing (ST6-OE) cells in CSC culture. Values = means and S.E. for 3 independent experiments, p<0.05; Student’s t test. (D) Immunoblot of SKOV3 cells cultured in CSC vs standard 10% FBS-containing media for 1 week. (E–F) ST6Gal-I immunoblotting and densitometry for SKOV3 (E) or MiaPaCa2 (F) cells cultured in CSC media for 1, 5, or 14 days. (G) ST6Gal-I expression (arrowhead) in ovarian ascites tumorspheroid. (H) SNA high (SNA-H) and SNA low (SNA-L) ascites cells grown in CSC culture. (I) Spheroids per field. (J) Ovarian PDX tumor SNA-H and SNA-L grown in CSC culture. (K) Viability of OV4 EV or ST6-OE cells incubated in cell-free ascites. Values = means and S.E., p<0.05; Student’s t-test. Scale bars=50µm.