JCS011262 Supplementary Material

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• Supplemental Figure S1 (Adobe PDF) -

  Fig. S1. TG catalytic activity of FXIIIA protein preparations. Recombinant FXIIIA proteins (0.1 µg) were analyzed for TG activity in the presence and absence of thrombin, as described in Materials and Methods. (n=9). *P<0.05 for mutant FXIIIA relative to wild-type FXIIIA under the same conditions with respect to presence or absence of thrombin.

• Supplemental Figure S2 (Adobe PDF) -

  Fig. S2. sFXIIIA has no effect on TG2 mRNA expression. Normal human knee articular chondrocytes plated in twelve-well dishes (at 1×10⁵ cells per well) and after a 2-hour incubation under serum-free conditions, the cells were incubated for 5 minutes with 100 ng/ml of recombinant FXIIIA in ascorbate-containing medium A, as described in the Materials and Methods. Using quantitative RT-PCR, we determined the TG2 chondrocyte mRNA expression levels relative to GAPDH, studying data from three separate human donors (n=9).

• Supplemental Figure S3 (Adobe PDF) -

  Fig. S3. Effects of FXIIIA mutants on p38 phosphorylation in chondrocytes. Aliquots of 1×10⁵ normal human knee articular chondrocytes were starved for 2 hours in serum-free DMEM and then incubated for the indicated times with 100 ng/ml of sFXIIIA recombinant proteins. The chondrocytes were examined for p38 phosphorylation by SDS-PAGE and western blotting. Representative of results from three experiments employing three separate donors.