RNAseq of ST6GAL1-wild type and ST6GAL1-knock out cells of MDA-MB-231 cells.

Dynamic change of glycosylation is found in various cancers; however, the precise role of glycosylation in metastasis is remained largely unknown. Current study show that α2,6-sialylation or β-galactoside α2,6-sialyltransferase (ST6Gal1) inhibits circulating tumor cell aggregation and depletion or restoration of α2,6-sialylation in patient derived xenograft model amplified or diminished cell aggregation and lung metastasis. ST6GAL1 was knocked out using CRISPR/Cas9 technology in MDA-MB-231 cells. To obtain differnt clones of ST6KO and ST6WT cells, single cells were sorted into 96 well/plate. Cells were checked using western blotting, and flow cytometry analysis. Cells were lysed for total RNA isolation using Trizol. Samples were sent to Northwestern University’s Center for Genetic Medicine Sequencing core facility for deep sequencing analysis.