We found 54 overexpressed and 24 underexpressed miRNAs with a FDR<15% in the KIT double mutated GISTs compared with the single mutated GISTs (Supplementary Table S8).
To determine whether KIT double mutated tumours differ from KIT single mutated tumours based on their miRNA expression levels and to identify the most significant deregulated miRNAs between these two groups, we performed SAM analysis on the samples harbouring only KIT mutations (four double KIT-mutated vs nine single KIT-mutated tumours) in the microarray data set.
Of specific relevance is the up-regulation of both VEGF-A [58] and VEGF-C [59] in response to over-expression or exogenous addition of either active or inactive (double mutated) heparanase.
Moreover, addition of recombinant, enzymatically inactive double mutated (DM) heparanase facilitated F cells adhesion similar in magnitude to the wild type protein (Fig. 1C&D, DM), further supporting the notion that improved cellular adhesion upon heparanase over-expression [27] or exogenous addition (Fig. 1) does not involve enzymatic aspects.
AGS cells were also co-transfected with the CDX2 expression vector or the corresponding empty vector, and the pCdx2-1luc wild-type, single and double mutated.
Female Tg2576 transgenic mice possessing a copy of human APP with the Swedish double mutate on (KM670/671NL) were obtained from Taconic and aged to 18 months.
While other cysteine residues affect antigenicity when mutated, a double mutation of these 121 124 cysteine residues has physical properties similar to those of wild-type virus (12 ).
Double-mutated, inactive heparanase (DM) and the KKDC peptide stimulated Rac1 activation to the same extent (Fig. 6A, DM, upper panel) or even higher (KKDC) than wild type heparanase (Fig. 6A, upper panel).
It also induces double mutations CC > TT, in which adjacent cytosines mutate to thymines as a result of cytosine dimers generated by UV light.
The AmVKRYYEE construct with a double mutation on K1043 and F1044 residues was obtained by mutating a selected clone of AmVKRYYEF with the 5'-CGAGAACGATTACTACGAGgagAATCGAAGAGGTATG-3' primer.
