Fig. 1

Selection of hCD39-specific ASO and assessment of target knockdown efficiency. (a) HDLM-2 cells were treated for three days with the indicated concentrations of the respective antisense oligonucleotide without the use of a transfection reagent. hCD39 mRNA expression values were normalized to expression of the housekeeping gene HPRT1. Residual hCD39 mRNA expression relative to untreated cells (set to 100%) is depicted. Depicted is the mean of triplicate wells +/− SD. (b) and (c) Human anti-CD3, anti-CD28, and IL-2 activated CD8⁺ and CD4⁺ T cells were treated with 5 μM of the hCD39-specific ASO A04040H or the control oligo 2 for a total treatment time of six days without the use of a transfection reagent. Thereafter, oligonucleotides were removed and cells were re-plated on plates not coated with anti-CD3. hCD39 protein expression was analyzed by flow cytometry three, six and eleven days after oligonucleotide removal. hCD39 protein expression is depicted as mean fluorescence intensity (MFI) and was calculated by subtracting the MFI of hCD39 by the MFI of unspecific isotype control (b) or as % CD39⁺ cells of alive cells (c). Data is shown as mean of duplicates +/−SD. Asterisks indicate significant differences compared to control oligo treatment within each time point