Fig. 1

OGT is required for the induction of EMT. A549 cells were transfected with OGT siRNA (+) or control siRNA (−) followed by culturing 3D spheroids with (+) or without (−) the addition of TNF and TGFβ. a Immunoblotting of total cell lysates demonstrates that OGT is required for the accumulation of O-GlcNAcylated proteins in cytokine-treated A549 cells. b RT-qPCR analysis confirms that OGT expression is required for the cytokine-induced expression of SNAI2, TWIST1 and ZEB2. Changes in mRNA expression were calculated relative to HPRT with mean and SD + shown; * p = < 0.05, ** = < 0.01, and *** < 0.005, n = 3. c Immunoblotting of total cell lysates demonstrates that OGT is required for cytokine-responsive upregulation of mesenchymal protein markers, N-cadherin, Fibronectin, and Vimentin. a & c Densitometic quantification of immunoblots demonstrates that the knockdown of OGT significantly dampens global O-GlcNAcylation and suppresses expression of mesenchymal protein markers relative to tubulin. Immunoblots are a representative example from at least three independent experiments