Hs stat3 7 flexitube sirna

HIF-1α siRNAs (Hs_HIF1α_5 FlexiTube siRNA, #SI02664053, Qiagen, Hilden, Germany) [64 (link)], scrambled (Scr) siRNAs (Negative Control siRNA, #1027310, Qiagen), STAT3 siRNAs (Hs_STAT3_7 FlexiTube siRNA, #SI02662338, Qiagen) [56 (link)] and Lipofectamine™ 2000 Transfection Reagent (Invitrogen, Carlsbad, CA, USA), were used to make complexes with a (siRNA:polymer) ratio of 1:1 (weight/weight) in OptiMEM media (Life Technologies, Grand Island, NY, USA) according to the manufacturer’s instructions. Complexes were added to the cells at 50 nM siRNA concentrations. Cells were transfected with siRNA complexes at 50–60% confluence.

Starting material for the preparation of peptide-based gadolinium nanoparticles was previously described [39 (link)]. Silencer^® Negative Control #1 siRNA, Catalog #: AM4635-AM4636 was purchased from Applied Biosystems, Ambian Inc., USA. Negative control siRNAs, the siRNAs with sequences that do not target any gene product—are essential for determining siRNA transfection efficiency and to control for the effects of siRNA delivery. We used this siRNA for performing binding affinity, DLS experiments, and flow cytometry experiments. In addition, Hs_STAT3_7 FlexiTube siRNA was purchased from QIAGEN LLC, USA. The target sequence was 5′-CAGCCTCTCTGCAGAATTCAA-3′, the sense strand was 5′-GCUUCUCUGCAGAAUUCAATT-3′ and the antisense strand was 5′-UUGAAUUCUGCAGAGAGGCTG-3′. STAT-3 (124H6) Mouse mAb #9139 and GAPDH (D4C6R) Mouse mAb #97166 were purchased from Cell Signaling Technology (CST), USA for the western blotting experiment. Horseradish peroxidase (HRP)-linked secondary antibody was purchased from Abcam Inc., USA. Mini-PROTEAN TGX stain-free precast gels and Trans-Blot Turbo Mini 0.2 µm PVDF transfer packs #1704156 were purchased from Bio-Rad Inc., Hercules, CA, USA. All other reagents used for western blotting experiments were purchased from Bio-Rad Inc., Hercules, CA, USA.