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. 2021 Mar 16;13(6):1330. doi: 10.3390/cancers13061330

Figure 3.

Figure 3

Characterization of biglycan expression in different GC cell lines and in knock out (KO) cells. (A) Biglycan expression in secretome samples from an intestinal cancer cell line Caco-2 (positive control) and from four GC cell lines (MKN74, MKN45, NCI.N87, and AGS) with and without chondroitinase ABC (chABC) treatment. Western blot analysis show MKN74 as the only GC cell line positive for biglycan. (B) Biglycan immunodetection in the MKN74 WT and KO cells (clones KO.5 and KO.12), showing the loss of expression in both KO clones. Chondroitinase-4-sulfate (C4S) immunedetection was used as a positive control for the chABC enzymatic treatment in all analysis. An increase in the C4S detection is expected after glycosaminoglycan (GAG) digestion by chABC. Ponceau staining was used as a loading control for secretome samples. (C) Bright field microscopy pictures (Magnification at 400×) and F-actin (red—phaloidin) immunofluorescence in MKN74 WT and biglycan KO cell (clones KO.5 and KO.12). Biglycan KO cells present morphological differences when compared with WT cells.