The spindle cell component stained positively, at least focally, for smooth muscle actin (data not shown).
Data presented thus far show calpain-1 within VSMC co-localizes with vimentin (Fig. 6A), is capable of cleaving spectrin [18], vimentin (Fig. 6B), and α-smooth muscle actin (data not shown), and is also linked to Ang II induced activation of MMP2.
A similar pattern of binding occurred following incubation with rabbit muscle actin (data not shown).
Downregulation of E-cadherin in our system was correlated with changes in tumour cell morphology but not with changes of vimentin or α-smooth muscle actin expression (data not shown).
We routinely immunostained comparably prepared cultures and, consistent with our prior report describing this method [24], found the majority of resultant cells to be comprised of cardiomyocytes (59±8% positive for the striated muscle marker sarcomeric actin, data not shown).
Stainings of the TMA with RMS markers, myogenin and desmin, was positive in both adherent cells and spheres (Figure 1E) which were negative for markers of other small blue round cell tumors (CD45, CD99, cytokeratin (CK), S100b, Synaptosin, smooth muscle actin (SMA) and WT1; data not shown).
In this study marked invasion into the connective tissue of the TEIOC model was evident even in the presence of normal human oral fibroblasts (NOFs were α-smooth muscle actin negative by immunohistochemistry; data not shown).
Changes in size of the spleen MGZ were analyzed after staining with CD20 (Figure 4.B), ASM (Alpha Smooth Muscle Actin) (Figure 4.C) or ICAM1 (data not shown) mAb.
As expected, TGF-β increased the number of cultured fibroblasts expressing α-smooth muscle actin as detected using immunofluoresence (data not shown).
Furthermore, immunohistochemical analyses revealed that the tumor cells were focally and weakly immunopositive for vimentin, and immunonegative for the differentiation markers cytokeratin, S100, desmin, α-smooth muscle actin, and neuron-specific enolase (data not shown).
Although mRNA expression of Sma, which encodes smooth muscle actin, was comparable between genotypes (data not shown), there was a trend towards increased Sma protein expression in LysM-Wls mice (Fig. 5d, e), suggesting an increase in myofibroblast activation in these mice.
