Transglutaminases Are Active in Perivascular Adipose Tissue

Figures

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  Figure 1. Expression of transglutaminase family genes in APVAT (blue), SMAPVAT (red), and MRPVAT (green). Multiple primers were used for TG1 and TG3, as indicated by set 1 and set 2. Boxes indicate target transglutaminases used for further studies. β-2-microglobulin was used as the reference gene. Data are expressed as mean ± SEM. n = 4 rats for each group.

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  Figure 2. Brightfield immunohistochemistry images using (A) TG2 or (B) FXIII primary antibodies to determine location of TGs in APVAT, SMAPVAT, and MRPVAT. Rat aorta and human placenta were used as positive controls for TG2 and FXIII, respectively. Arrows indicate locations of positive TG staining. Images are representative of n = 4.

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  Figure 3. Images of in situ activity signal using (A) TG2 T26 or (B) FXIII F11KA substrate peptides to determine TG activity in fresh frozen tissue sections. Red staining and arrows indicate locations of TG activity. Rat aorta and skin were used as positive controls for TG2 and FXIII, respectively. Images are representative n ≥ 4.

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  Figure 4. BAP incorporation into MRPVAT, visualized with IRDye 800CW Streptavidin secondary (A top) and total protein stain (A bottom). Quantification of the relative signal normalized to the total protein stain (B). Data are expressed as mean ± SEM. n = 7 rats per group. **** p < 0.0001 compared to only CaCl2 treated MRPVAT protein by one-way ANOVA with Bonferroni’s multiple comparison.

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  Figure 5. BAP incorporation into MRPVAT adipocytes and SVF visualized with IRDye 800CW Streptavidin secondary (A top) and total protein stain (A bottom). Quantification of the relative signal normalized to the total protein stain (B). Data are expressed as mean ± SEM. n = 3 rats per group for adipocyte samples and n = 4 rats per group for SVF samples. ** p < 0.01 compared to only CaCl2 treated MRPVAT adipocyte protein by one-way ANOVA with Bonferroni’s multiple comparison.

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  Figure 6. BAP incorporation into MRPVAT adipocytes in the presence of increasing NE concentrations (mM), denoted by numbers below gradient triangles (A), visualized with IRDye 800CW Streptavidin secondary (A top) and total protein stain (A bottom). Quantification of the relative signal normalized to the total protein stain (B). Data are expressed as mean ± SEM. n = 4 rats per group. ** p < 0.01, * p < 0.05 compared to only CaCl2 treated MRPVAT adipocyte protein by one-way ANOVA with Bonferroni’s multiple comparison.