Figure 4.

TLC and cell cycle determination of UGCG-gRNA treated HCT116- and Lovo cells. (A,D) An almost complete depletion of neutral and acidic GSLs was achieved in HCT116- (A) and Lovo cells (D) by UGCG-guide-RNA (gRNA); x, no GSL-positive band; running solvent for neutral- and acidic GSLs was CHCl₃/CH₃OH, 0.2% CaCl₂, 60:35:8 (v/v). (B,C, quantification) UGCG-gRNA treated HCT116 cells did not show an arrest of the cell cycle, similar as HCT116 cells treated with low doses of Genz (Figure 2). (E–G, quantification) Genetic depletion using UGCG-gRNA-application (E) and treatment of Lovo cells with 5 µM Genz (F) resulted in an arrest of the cell cycle, whereas the response in Genz-treated cells was slightly more pronounced; significances are *, p ≤ 0.05; **, p < 0.01; ***, p < 0.001.