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. 2021 Feb 12;11:3718. doi: 10.1038/s41598-021-82686-3

Figure 4.

Figure 4

Anti-Gal3 monoclonal antibody, 14D11, inhibits MUC16-mediated tumorigenesis in vitro. (a) Direct ELISA for Gal3 binding to selected antibodies in the presence or absence of lactose. The presence of increasing lactose concentrations progressively inhibited the binding of Gal3 to 14D11, an isotype control antibody (18C6) and 1F5, an antibody reactive to the N terminal 15 amino acids of Gal3. Each lactose condition was compared to the zero lactose condition for the same antibody by paired t-test. (b) Effect of 14D11 on MUC16-mediated activation of ERK/AKT signaling in MUC16-expressing A2780c344 and SKOV3c344 cells after 48-h exposure to antibody. MUC16-negative A2780 and SKOV3 cells were used as controls. Densitometry was used to normalize the results against β-actin. (c) Effect of 14D11 on Matrigel invasion in MUC16-overexpressing SKOV3c344 and A2780c344 cells, and endogenous MUC16-expressing OVCAR3 cells. Anti-MUC16 N-glycosylation site antibody, 18C6, was used as a positive control. The values are the mean of triplicate well from a representative study and the invasion assays were repeated independently ≥ 3times. (d) Effect of 14D11 on Matrigel invasion in MDA-MB-231 and shLGALS3-MDA-MB-231 cells, performed as in 3C.