Figure 1.

Optimization of the transfection efficiency of patient-derived childhood ALL cells. A) ALL cells of patient ALL-50 were xenografted and amplified in NOD/SCID mice, isolated from spleens and transfected by nucleofection with a control siRNA oligonucleotide conjugated to Alexa-Fluor-488 using different programs. After 48 h of in vitro culture, apoptosis was measured by forward side scatter analysis and transfection efficiency in living cells by the shift of fluorescence intensity in channel 1 in FACscan. Presented are original results of 4 different transfection conditions for sample ALL-50. B) The data of eight different transfection conditions (X-Axis indicates the different programs offered by the company) are summarized for transfection efficiency and impact of transfection on survival of ALL cells for sample ALL-50. * highlights the program used for further experiments. C) Absolute cell count was determined in ALL-50 cells untreated (co) and transfected with the dye-conjugated control sequence (sicontrol) at 0 and 48 h. D) Transfection efficiency and cell viability was determined in overall n = 11 different xenograft samples using program C16 as in Figure 1B. E) ALL cells from patient ALL-50 were simultaneously transfected with a control siRNA against Lamin conjugated to Alexa-Fluor-488 and siRNA control oligonucleotide conjugated to Alexa-Fluor647 as in Figure 1B using program C16.