One of the most investigated housekeeping proteins is Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which plays an important role across a diverse array of biological functions. GAPDH is widely used as an internal control in analysis of gene and protein expression levels. It is also a target of EHEC NleB1, and its glycosylation inhibits GAPDH interaction with TRAF2, which contributes to inhibiting NF-κB pathway activation. . Kansas State University researchers developed a GAPDH knockdown stable cell line to show that NleB/SseK orthologs have different acceptor substrate specificities. ShRNAs were synthesized and cloned into a mammalian expression vector. The plasmid was transfected into HEK 293T cells. Stable cell lines were created using puromycin selection. Caronda Moore caronda@ksu.edu 785-532-1366
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