Validation of AS1 as novel regulator of tumor resistance to immune cell attack

The experiment was performed to verify wether knockdown of AS1 in combination with TNFa treatment affected the signature of genes involved in tumor cell apoptosis and survival Overall design: The experiment was conducted with biological replicates. PANC-1 WT cells were reverse transfected either with siCtr1 or with siAS1 s1 (siRNA sequences) in a 6-well plate . Afterwards, the medium was replaced with 1 mL of complete DMEM and 1 mL rHuTNF-a was added at a final concentration of 100 ng/mL. After 30 min or 4h from stimulation, tumor cells were harvested in trypsin and RNA was isolated using the RNeasy mini kit (Qiagen) according to the manufacturer´s instructions.