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. 2021 May 13;22(10):5144. doi: 10.3390/ijms22105144

Figure 4.

Figure 4

HUVECs after incubation in a cultivation medium containing Gal-3 (30 μM) or LacdiNAc (1 mM) or both molecules added together or in a different order. Control cells were incubated for 1 h in a pure cultivation medium. (A) Intensity of the fluorescence signal in HUVECs stained for Gal-3 (ex./em. = 485/528 nm). The data are shown as the mean ± SD from 3 wells. Holm-Sidak test, p ≤ 0.05. * Statistically significant difference in comparison with cells in the pure medium (ctrl), # Statistically significant difference in comparison with cells incubated with Gal-3 (Gal-3). (B) Immunofluorescence of Gal-3 in the cells compared in graph A. Anti-Gal-3 antibody (EP2775Y) produced in rabbits (Abcam, Cambridge, UK; Cat. No. ab76245) was used. The cell nuclei were counterstained with Hoechst 33258. Olympus IX 71 microscope, DP 70 digital camera, obj. 20×, scale bar 50 μm. Ctrl: cells in the pure medium; Gal3: cells incubated for 1 h in the medium with Gal-3; LacdiNAc: cells incubated for 1 h in the medium with 1-mM LacdiNAc; Gal3 + LacdiNAc: cells incubated for 1 h in the mixture of Gal-3 and LacdiNAc; LacdiNAc, Gal3: cells incubated for 1 h in the medium with LacdiNAc, then for 1 h in the medium with Gal-3; Gal3, LacdiNAc: cells incubated for 1 h in the medium with Gal-3, then for 1 h in the medium with LacdiNAc.