Figure 6

Impact of Msp1 glycosylation on Akt activation in intestinal epithelial cells. Caco-2 cells were co-incubated with L. rhamnosus GG wild-type (10⁷ CFU/ml), msp1 mutant CMPG10200 (10⁷ CFU/ml), glycosylated Msp1 after purification with affinity chromatography and ConA sepharose (ca. 100 ng/ml), Msp1 partially purified from spent culture supernatant (SN) (ca. 10 ng/ml), chemically deglycosylated Msp1 after TFMS treatment (ca. 100 ng/ml) and recombinantly (rec.) expressed Msp1 after purification from E. coli (ca. 100 ng/ml). Subsequent Akt activation was monitored by Western blot analysis of cellular lysates with anti-P-Akt and ant-Akt antibodies. Data are representative for 2 separate experiments.