Fig. 2

Glyoxalase 1 (GLO1) depletion modifies the expression of extracellular matrix (ECM) proteins by breast cancer cells and promotes collagen gel retraction and anchorage-independent growth. a Collagen (COL)4A3, COL4A4, COL6A1, COL6A2 and COL6A3 messenger RNA (mRNA) levels were assessed by qRT-PCR in GLO1-depleted MDA-MB-231 cells. Data were analyzed using one-way analysis of variance (ANOVA) followed by Dunnett post-hoc test and shown as mean values ± SEM of three independent experiments. b Collagen VI α1 and α3 protein levels in MDA-MB-231 shGLO1 cells. Immunoblot data were quantified by densitometric analysis and normalized to Hsc70. Numbers represent fold increase relatively to the condition shown with bold number. Western blots are representative of two independent experiments. c Collagen VI α1 and α3 protein levels in MDA-MB-231 shGLO1#2 and control (shNT) cells were assessed by immunohistochemical analysis (IHC). d Representative collagen gel retraction assays performed with MDA-MB-231 shGLO1 cells. e Quantification of collagen gel area after 6 days. Data were analyzed using one-way ANOVA followed by Dunnett post-hoc test and shown as mean values ± SEM of five independent experiments. f Quantification of colonies formed in a soft agar matrix by GLO1-silenced MDA-MB-231 cells treated with carnosine. Data were analyzed using two-way ANOVA followed by Bonferroni post-hoc test and shown as mean values ± SEM of three independent experiments. ns, not significant; *p < 0.05, **p < 0.01 and ***p < 0.001