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. 2020 Dec 12;70(10):1872–1883. doi: 10.1136/gutjnl-2020-322468

Figure 4.

Figure 4

Bacterial secreted OGAs can hydrolyse O-GlcNAcylated proteins in human enterocyte-like Caco-2 cells. (A) Schematic of BtGH84 and AkkGH84 domain organisation, the active catalytic sites were indicated in red. (B) Polyclonal antibody against BtGH84 or AkkGH84 was generated and used to detect OGAs in bacterial lysates and supernatant by immunoblot analysis. (C, D) Immunoblot analysis of O-GlcNAc levels in Caco-2 lysates, after incubating the cell lysates with different amounts of BtGH84 or AkkGH84 (C) or their active sites mutants BtGH84-2D or AkkGH84-2D (D). (E, F) Immunoblot analysis of O-GlcNAc levels in Caco-2, after adding into the cell culture supernatant with different amounts of BtGH84 or AkkGH84 (E) or the Thiament G, an OGA inhibitor (F). Data represent the mean of three repeats per group with the SD; **P<0.01, ***P<0.001. OGA, O-GlcNAcase.