Silencer select pre designed validated sirna

Gene-knockdown of CXorf21 in LCLs (GM12878) was performed by siRNA using the Nucleofector II Device (Lonza) and Amaxa Cell Line Nucleofector Kit V. Two days before transfection, cells were seeded to a concentration of 0.5 × 10⁶ cells/ml. In duplicate, 2 × 10⁶ cells were spun at 100 g for 10 min and re-suspended in 100 μl supplemented transfection solution and 20 pmol Silencer Select Pre-Designed & Validated siRNA (Thermo Fisher Scientific) against CXorf21 (#4392420). The Silencer Select Negative Control No. 1 siRNA (#4390843) was used as a non-targeting negative control at the same concentration. Cell/siRNA suspensions were transferred to a Nucleofector cuvette and electroporated using the X-001 programme. Samples were cultured in 1.5 ml medium in a 12-well plate and harvested 48 h post-transfection.

ALPP siRNA (Silencer™ Select Pre-designed & Validated siRNA; s1300 and s1301, Thermo Fisher Scientific) and negative control siRNA (Silencer™ Select Negative Control No. 1 siRNA, Thermo Fisher Scientific) were added 1:1 to Lipofectamine RNAiMAX (Thermo Fisher Scientific). MDA-MB-231 was seeded into 6-well plates at a cell density of 2.0 × 10⁵ cells/ml for 3 days. Cells were cultured after addition of OC, OC with siRNA 10 µM (s1300 + s1301), or OC with negative control siRNA 10 µM. We performed western blot after 2 weeks and alizarin and ALP staining after 6 weeks.