Fig. 2.

GRASP KO increases HS synthesis but reduces its sulfation and secretion. A GRASP KO increases HS synthesis analyzed by LC–MS. Shown are the total amount of HS in both cells and medium of indicated cell lines. B GRASP KO increases HS synthesis analyzed by immunofluorescence microscopy. Indicated cells were permeabilized and stained for HS with an HS antibody 10E4. Shown are microscopic images (left) and quantitation (right). C GRASP KO increases HS synthesis analyzed by flow cytometry. Indicated cells were permeabilized, stained for HS with an HS antibody and analyzed by flow cytometry. D GRASP KO decreases HS secretion. The percentage of secreted HS (HS in conditioned media/HS in both cell lysate and conditioned media) was analyzed by LC–MS. E GRASP KO reduces HS sulfation in the cell lysate. Shown is the percentage of each sulfated form of HS in cells. F GRASP KO reduces HS sulfation in the conditioned media. Shown is the percentage of each sulfated form of HS in the conditioned media