Fig. 2
From: Identification of O-GlcNAcylated proteins in Plasmodium falciparum
A large panel of proteins expressed by P. falciparum are O-GlcNAcylated. P. falciparum proteins were analysed by western blot using the anti-O-GlcNAc antibody RL2 (a) or after labelling of the O-GlcNAc-bearing proteins by GalNAz and biotin alkyne using the Click-it™ O-GlcNAc enzymatic labelling system and the Click-it™ Glycoprotein detection kit as described in the “Methods” section (b). After labelling P. falciparum O-GlcNAcylated proteins were separated by SDS-PAGE and western blot analyses were performed using HRP-labelled avidin to test the efficiency of the labelling (c). After enzymatic and chemical labelling, P. falciparum O-GlcNAcylated proteins were enriched on avidin-beads, separated by SDS-PAGE, electroblotted onto nitrocellulose membrane and analysed by western blot according to their α-tubulin and Hsp70 contents. The clicked-proteins were enriched on avidin-beads and analysed by Western blot by according to their alpha-tubulin and Hsp70 antibodies (d)