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. 2021 May 25;10(6):1310. doi: 10.3390/cells10061310

Figure 1.

Figure 1

UEAI recognizes intracellular proteins. (A) WM793 cells were fixed and permeabilized with Triton X-100 or left unpermeabilized. Cells were subsequently stained with DAPI nuclear stain, FITC-AAL, FITC-UEAI, or no lectin as a control. Cell were visualized by IF for qualitative lectin staining (left panel). For quantification, WM793 and A375 cells were stained as above and analyzed by flow cytometry (right panel; MFI: Mean Fluorescence Intensity (of UEA1 per cell)). * p = 0.006 (B) Cells transduced and selected with lentivirus containing Cas9/no guide (ng) or Cas9/sgSLC35C1 (gC1) were grown in normal growth media and harvested for protein lysate. Lysates were probed with AAL and UEAI lectin to determine global levels of Golgi-dependent fucosylation. (C) WM793 cells were cultured for three days in 2-fluorofucose (2-FF, FUTi, 250 µM), L-fucose (L-fuc, 250 µM), or control. Cells were harvested for protein lysate and blotted for global AAL and UEAI levels.