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. 2021 Jul 29;7:197. doi: 10.1038/s41420-021-00543-8

Fig. 3. TG2 crosslinking function is lost but GTP, integrin, and fibronectin interactions are preserved in Tgm2-C277S mice.

Fig. 3

A (i) Representative western blot and densitometry analysis showing that TG2 expression levels in aortic homogenates of Tgm2-C277S heterozygous (HET) and homozygous (HOM) mice are similar to those in littermate WT mice. Age-matched TG2−/− mice were used as controls (n = 8 per group). TG2 activity was significantly lower in Tgm2-C277S mice, as measured by (ii) crosslinking of FITC-cadaverine and N,N′-dimethylcasein determined using fluorescence polarization (FP) and (iii) biotin(amido)pentylamine (BPA) incorporation in intact aortic rings by dot blotting in the presence of DTT (n = 8 per group; *p < 0.05, **p < 0.01, ***p < 0.001 by one-way ANOVA with Bonferroni post hoc analysis). (iv) Representative en face confocal microscopy images show TG2-dependent incorporation of FITC-cadaverine into the aortic media of Tgm2-C277S mice and WT littermates in the presence of DTT, a TG2 activator. Age-matched TG2−/− (KO) mice were used as controls. Blue = DAPI; green = FITC-cadaverine; n = 5 per group. B Representative western blots of WT and Tgm2-C277S mouse liver show GTP binding of TG2; TG2−/− mice were used as controls. C Representative western blots of WT and Tgm2-C277S mouse liver show co-immunoprecipitation of TG2 with integrin β1 and fibronectin and vice versa. Blots showing total expression of these proteins are shown for reference. (n = 5 mice per group; **p < 0.01; ****p < 0.0001 by ordinary one-way ANOVA with Bonferroni post hoc analysis).