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. Author manuscript; available in PMC: 2022 Jun 4.
Published in final edited form as: Cell Mol Life Sci. 2022 Mar 21;79(4):199. doi: 10.1007/s00018-022-04223-3

Figure 3. GRASP KO reduces CS synthesis and secretion.

Figure 3.

(A) GRASP KO reduces CS synthesis analyzed by LC-MS. Shown are the total amount of CS in both cells and medium of indicated cell lines. (B) GRASP KO decreases CS synthesis analyzed by immunofluorescence microscopy. Indicated cells were permeabilized and stained for CS with a CS antibody (CS-56). Shown are microscopic images (left) and quantitation (right). (C) GRASP KO decreases CS synthesis shown by flow cytometry. Indicated cells were permeabilized, stained for CS with a CS antibody and analyzed by flow cytometry. (D) GRASP KO decreases CS secretion. The percentage of secreted CS (CS in conditioned media/CS in both cell lysate and conditioned media) was analyzed by LC-MS. (E) GRASP KO alters CS sulfation in the cell lysate. Shown are the percentage of each sulfated form of CS in cells. (F) GRASP KO alters CS sulfation in the conditioned media. Shown are the percentage of each sulfated form of CS in the conditioned media. Note that GRASP depletion increased 4-sulfation while decreases 6-sulfation in both the cell lysate (E) and conditioned media (F).