Figure 6: Depletion of GRASP results in altered cell surface glycosylation.
From: Regulation of protein glycosylation and sorting by the Golgi matrix proteins GRASP55/65
(a–d) Surface staining of fluorophore-conjugated lectin MAA specific for α(2,3) SA was reduced upon the depletion of GRASPs. Non-permeabilized HeLa cells treated with the indicated RNAi were exposed to tetramethylrhodamine isothiocyanate-conjugated MAA. The cells were then fixed, stained with GRASP antibodies and visualized using a confocal microscope. Scale bar, 50 μm. (e–f) HeLa cells transfected with control RNAi (arrows) and with RNAi for GRASP65 and 55 (asterisks) were mixed, cultured together and stained for GRASP65 (green) and WGA (red). Scale bar, 20 μm. (g) GRASP depletion impacts Lamp1 and Lamp2 glycosylation. Indicated GRASP-depleted cells were analysed for Lamp1, Lamp2 and p97 by western blots. Molecular weight markers are shown on the right. Note the shift of the bands after GRASP depletion.
