Skip to main content
. 2021 May 25;9:659428. doi: 10.3389/fcell.2021.659428

FIGURE 1.

FIGURE 1

C4ST-1 deficiency suppresses the proliferation of BT-549 cells. (A) Growth curves of parental BT-549 and C4ST-1 KO cells. (B) Proliferation of BT-549 (n = 4) and C4ST-1 KO cells (n = 4) was measured by CytoTox-ONETM Assay. (C) Proliferation of BT-549 and C4ST-1 KO cells was examined by colony formation assay. Both the cell types were seeded at a concentration of 50 cells/well in 6-well plate and cultured for 9 days. The colonies were stained with crystal violet, and observed under a light microscope (Left). The number of colonies of BT-549 (n = 3) and C4ST-1 KO cells (n = 3) was compared (Right). (D) The level of cyclin D1 in BT-549 and C4ST-1 KO cells was measured by real-time PCR (n = 3 each). (E) Proliferation of BT-549 (n = 4) and C4ST-1 KO cells (n = 4) treated with or without Chase ABC was examined by CytoTox-ONETM Assay. Cells were digested in the serum-free medium ASF Medium 104 for 4 days by adding 5 munits/well of Chase ABC twice at 0 and 2 days. (F) Cells digested with or without Chase ABC were stained by anti-CS antibody (clone 2B6), which detects the terminal unsaturated disaccharide of CS chains generated by Chase ABC. (G) Proliferation of BT-549 (n = 4) and C4ST-1 KO cells (n = 4) treated with or without GM6001 was measured. Statistical significance was determined using Student’s t-test. Statistical analyses were performed using KaleidaGraph version 4.5.1.